MXPA99009327A - Methods and compositions for synthesis of long chain polyunsaturated fatty acids - Google Patents

Abstract

The present invention relates to a fatty acid&Dgr;5-desaturase able to catalyze the conversion of dihomo-gamma-linolenic acid to arachidonic acid. Nucleic acid sequences encoding&Dgr;5-desaturase, nucleic acid sequences which hybridize thereto, DNA constructs comprising a&Dgr;5-desaturase gene, and recombinant host microorganism or animal expressing increased levels of a&Dgr;5-desaturase are described. Methods for desaturating a fatty acid at the&Dgr;5 position and for producing arachidonic acid by expressing increased levels of a&Dgr;5-desaturase are disclosed. Fatty acids, and oils containing them, which have been desaturated by a&Dgr;5-desaturase produced by recombinant host microorganisms or animals are provided. Pharmaceutical compositions,infant formulas or dietary supplements containing fatty acids which have been desaturated by a&Dgr;5-desaturase produced by a recombinant host microorganism or animal also are described.

Description

METHODS AND COMPOSITIONS FOR THE LONG CHAIN POLY-INSATURED FATTY ATS S NTESIS INTRODUCTION Field of the Invention This invention relates to modulating levels of enzymes and / or enzyme components that are related to the production of long-chain polyunsaturated fatty acids (PUFA) in a micro-organism or animal. Background Two main families of polyunsaturated fatty acids are the fatty acids? 3, exemplified by eicosapentaenoic acid (AEP), and? 6 fatty acids, exemplified by arachidonic acid (ARA). Polyunsaturated fatty acids are important components of the plasma membrane of the cell, where they can be found in forms such as phospholipids. Polyunsaturated fatty acids are necessary for proper development, particularly in the development of the infant brain, and for the formation and repair of tissues. Unsaturated fatty acids also serve as precursors to other molecules of importance in humans and animals, including prostacyclines, eicosanoids, leukotrienes and prostaglandins. Four major long-chain polyunsaturated fatty acids of importance include docosahexaenoic acid (ADH) and eicosapentaenoic acid, which are mainly found in different types of fish oil, gamalinolenic acid (GLA), which is found in the seeds of several plants, including primrose (Oenothera biennis), borage (Borago officinalis) and black currant (Ribes nigrum), and stearidonic acid (AED), which is found in marine oils and plant seeds. Both the gamalinolenic acid and another important long-chain polyunsaturated fatty acid, arachidonic acid, are found in filamentous fungi. Arachidonic acid can be purified from animal tissues including the liver and the adrenal gland. Gamalinolenic acid, arachidonic acid, eicosapentaenoic acid and stearidonic acid are themselves, or are dietary precursors, important long chain fatty acids involved in the synthesis of prostaglandin, in the treatment of heart disease, and in the development of tissue gone brain. Polyunsaturated fatty acids have several pharmaceutical and medical applications that include the treatment of heart disease, cancer and arthritis. For docosahexaenoic acid, there are several sources for commercial production that include a variety of marine organisms, oils obtained from cold seawater fish, and egg yolk fractions. For arachidonic acid, micro-organisms that include the genera Mortierella, Entomophthora, Phytium and Porphyridium can be used for commercial production. Commercial sources of stearidonic acid include the genera Trichodesma and Echium. Commercial sources of gamalinolenic acid include primrose, black currant and borage. However, there are several disadvantages associated with the commercial production of polyunsaturated fatty acids from natural sources. Natural sources of polyunsaturated fatty acids, such as animals and plants, tend to have very heterogeneous fat compositions. The fats obtained from these sources may therefore require much purification to separate one or more desired polyunsaturated fatty acids or to produce a fat that is enriched in one or more polyunsaturated fatty acids. Natural sources are also subject to uncontrollable fluctuations in availability. Fish stocks may undergo natural variation or may be depleted by overfishing. Fish fats have unpleasant tastes and odors, which may be impossible to economically separate from the desired product, and may render these products unacceptable as food supplements. Animal fats, and particularly fish fats, can accumulate environmental pollutants. Climate and diseases can cause fluctuations in the performance of both fish and plant sources. The crop land available for the production of alternative crops that produce oils is subject to competition from the stable expansion of human populations and the associated growing need for food production in the rest of the arable land. Cultures that produce polyunsaturated fatty acids, such as borage, have not been adapted to the commercial crop and may not have good yield in monoculture. The growth of these crops is not economically competitive when other more profitable and better established crops can be grown. Large-scale fermentation of organisms such as? Tortierella is also expensive. Natural animal tissues contain low amounts of arachidonic acid and are difficult to process. Micro-organisms such as Porphyridium and Mortierella are difficult to grow on a commercial scale. Dietary supplements and pharmaceutical formulations containing polyunsaturated fatty acids may retain the disadvantages of the source of the polyunsaturated fatty acid. Supplements such as fish oil capsules may contain low levels of the particular component desired and thus require large doses. High doses result in the ingestion of high levels of unwanted components, including contaminants. The unpleasant flavors and odors of the supplements may make these regimens undesirable, and may inhibit patient cooperation. Care must be taken in providing supplements with fatty acids, as it can result in over-addition in the suppression of endogenous biosynthetic pathways and lead to competition with other fatty acids needed in various lipid fractions in vivo, leading to undesirable results. For example, Eskimos who have a diet high in fatty acids? 3 have an increasing tendency to bleed (Patent of the United States of America number 4,874,603). Several enzymes are involved in the biosynthesis of polyunsaturated fatty acids. Linolenic acid (LA, 18: 2 → 9, 12) is produced from oleic acid (18: 1 → 9) by an α2-desaturase. The gamalinolenic acid (18: 3? 6, 9, 12) is produced from linoleic acid (LA, 18: 2? 9, 12) by a? 6-desaturase. The production of arachidonic acid (20: 4? 5, 8, 11, 14) is produced from dihomo-gamma-linolenic acid (ADGL, 20: 3? 8, 11, 14) is catalyzed by a? 5-desaturase . However, animals can not desaturate beyond position? 9 and therefore can not convert oleic acid (18: 1? 9) into linolenic acid (18: 2? 9, 12). In the same way, linolenic acid (ALA, 18: 3? 9, 12, 15) can not be synthesized by mammals. Other eukaryotes, including fungi and plants, have enzymes that desaturate at the? 12 and? 15 positions. The polyunsaturated fatty acids greater than animals are therefore derived either from the diet and / or from the desaturation and elongation of linoleic acid (18: 2? 9, 12) or of c-linolenic acid (18: 3? 9, 12, 15). It is therefore interesting to obtain genetic material involved in the biosynthesis of polyunsaturated fatty acids from species that naturally produce these fatty acids and express the isolated material in a microbial or animal system which can be manipulated to provide the production of of one or more polyunsaturated fatty acids. Thus there is a need for desaturases of fatty acids, genes that encode them, and recombinant methods to produce them. There is also a need for fats containing relatively higher proportions and / or enriched in specific polyunsaturated fatty acids. There is also a need for reliable economic methods of production of specific polyunsaturated fatty acids. Relevant Literature The production of gamma-linolenic acid by a? 6-desaturase is described in the patent of the United States of North America Number: 5,552,306. The production of 8, 11-eicosadienoic acid using Mortierella alpina is described in U.S. Patent Number: 5,376,541. The production of docosahexaenoic acid by dinoflagellates is described in U.S. Patent Number: 5,407,957. The cloning of a carrier protein-6-palmitoyl-acyl desaturase is described in the publication of TCP WO 96/13591 and in the patent of the United States of America number: 5,614,400. The cloning of a? 6-desaturase from borage is described in the publication of TCP WO 96/21022. The cloning of? 9-desaturases is described in the published patent applications of TCP WO 91/13972, EP 0 550 162 Al, EP 0 561 569 A2, EP 0 644 263 A2, and EP 0 736 598 A1, and in the Patent of the United States of America Number: 5,057,419. The cloning of β2-desaturases from various organisms is described in the publication of TCP WO 94/11516 and the patent of the United States of America number: 5,443,974. The cloning of α5-desaturases from various organisms is described in the publication of TCP WO 93/11245. All publications and patents of the United States of North America or applications referred to herein are hereby incorporated by reference in their entirety. Summary of the Invention [0002] Novel compositions and methods for the preparation of polyunsaturated long chain fatty acids are provided. The compositions include nucleic acids encoding a? 5-desaturase and / or polypeptides having? 5-desaturase activity, the polypeptides, and the probes for isolating and detecting them. The methods involve culturing a host-organism or host animal that contains and expresses one or more transgenes encoding a? -desaturase and / or a polypeptide having? 5-desaturase activity. Expression of the desaturase polypeptide provides a relative increase in the 5-unsaturated polyunsaturated fatty acid, or in the metabolic progeny thereof, as a result of altered concentrations of enzymes and substrates involved in the biosynthesis of polyunsaturated fatty acid. The invention can be used for example in the large-scale production of polyunsaturated fatty acids containing oils including, for example, arachidonic acid, eicosapentaenoic acid and / or docosahexaenoic acid. In a preferred embodiment, a nucleic acid sequence comprising a 5-desaturase represented in Figure 3A-D (SEQ ID NO: 1), a polypeptide encoded by the nucleic acid, and a purified or isolated polypeptide is provided. in Figure 3A-D (SEQ ID NO: 2), and an isolated nucleic acid encoding the polypeptide of Figure 3A-D (SEQ ID NO: 2). Another embodiment of the invention is an isolated nucleic acid sequence encoding a polypeptide, wherein the polypeptide desaturates a fatty acid molecule at the carboxyl end carbon of the molecule. The nucleic acid is preferably derived from a eukaryotic cell, such as a fungus cell, or a fungus cell of the genus Mortierella, or from the genus / species Mortierella alpina. Also preferred is an isolated nucleic acid comprising a sequence that is attached to a nucleotide sequence depicted in Figure 3A-3D (SEQ ID NO: 1), and a nucleic acid encoding an amino acid sequence depicted in Figure 3A- D (SEQ ID NO: 2). In particular, the nucleic acid encodes an amino acid sequence represented in Figure 3A-D (SEQ ID NO: 2); the sual is selected from the group consisting of amino acid residues 30-38, 41-44, 171- 175, 203-212, and 387-394. In a further embodiment, the invention provides an isolated or purified polypeptide that desaturates a fatty acid molecule at the carbon 5 of the carboxyl terminus of the molecule. Also provided is an isolated nucleic acid sequence that hybridizes to a nucleotide sequence depicted in Figure 3A-D (SEQ ID NO: 1), an isolated nucleic acid sequence having at least 50% identity to the Figure 3A-D (SEQ ID NO: 1). The present invention further includes a nucleic acid construct comprising a nucleotide sequence depicted in Figure 3A-D (SEQ ID NO: 1) linked to a heterologous nucleic acid.; a nucleic acid construct comprising a nucleotide sequence depicted in Figure 3A-D (SEQ ID NO: 1) operably linked to a promoter; and a nucleic acid construct comprising a nucleotide sequence depicted in Figure 3A-D (SEQ ID NO: 1) operably linked to a promoter which is functional in a microbe cell. In a preferred embodiment, the microbe cell is a yeast cell, and the nucleotide sequence is derived from a fungus, such as a fungus of the genus Mortierella, particularly a fungus of the species Mortierella alpina. In another embodiment of the invention, there is provided a nucleic acid construct comprising a nucleotide sequence encoding a polypeptide comprising an amino acid sequence that corresponds to or is complementary to an amino acid sequence depicted in Figure 3A-D (SEQ ID NO: 2), wherein the nucleotide sequence is operably linked to a promoter that is functional in a host cell, and wherein the nucleotide sequence encodes a polypeptide that desaturates a fatty acid molecule at carbon 5 of the carboxyl end of the fatty acid molecule. Additionally, a nucleic acid construct comprising a nucleotide sequence encoding a functionally active? 5-desaturase is provided by the invention, wherein the desaturase includes an amino acid sequence that corresponds to or is complementary to all or a portion of a sequence of amino acids represented in Figure 3A-D (SEQ ID NO: 2), wherein the nucleotide sequence is operably linked to a functional promoter in a host cell. The invention also includes a host cell comprising a nucleic acid construct of the invention. In a preferred embodiment, there is provided a recombinant host cell comprising at least one copy of a DNA sequence encoding a functionally active Mortierella alpina fatty acid desaturase having an amino acid sequence as depicted in Figure 3A-D ( SEQ ID NO: 2), wherein the cell or an ancestor of the cell was transformed with a vector comprising that DNA sequence, and wherein the DNA sequence is operably linked to a promoter. The host cell may be eukaryotic or prokaryotic. Preferred eukaryotic host cells are those selected from the group consisting of a mammalian cell, an insect cell, a fungal cell, and an algal cell. Preferred animal cells include a bird cell, a fungus cell such as a yeast, and a seaweed cell. Preferred prokaryotic cells include those selected from the group consisting of a bacterium, a cyanobacterium, cells that contain a bacteriophage, and / or a virus. The DNA sequence of the recombinant host cell preferably contains a promoter that is functional in the host cell. The host cells of the invention containing the DNA sequences of the invention are enriched for fatty acids, such as 20: 3 fatty acids. In a preferred embodiment, the host cells are enriched by 20: 4-fatty acids compared to an untransformed host cell lacking said DNA sequence, and / or enriched by 20: 5 fatty acids compared to a non-host cell. transformed that lacks said DNA sequence. In another preferred embodiment, the invention provides a recombinant host cell comprising a fatty acid selected from the group consisting of a dihomo-α-linolenic acid, n-6 eicosatrienoic acid, 20: 3n-6 acid and acid : 3 (8, 11, 14). The present invention also includes a method for the production of arachidonic acid in a cell culture. < microbial, wherein the method comprises culturing a microbial cell culture having a plurality of microbial cells containing one or more nucleic acids encoding a polypeptide which converts the dihomo-γ-linolenic acid into arachidonic acid, wherein the nucleic acid it is operably linked to a promoter, under conditions whereby the one or more nucleic acids are expressed, whereby the arachidonic acid is produced in the microbial cell culture. In various preferred embodiments of the invention, the polypeptide is an enzyme that desaturates a fatty acid molecule at carbon 5 from the carboxyl terminus of the fatty acid molecule; the nucleic acid is derived from a Mortierella sp.; and the substrate for the polypeptide is supplied exogenously. The microbial cells used in the methods can be either eukaryotic cells or prokaryotic cells. Preferred eukaryotic cells are those selected from the group consisting of a mammalian cell, an insect cell, a fungus cell, and an algae cell. Preferred animal cells include a bird cell, a preferred fungal cell is a yeast, a preferred algae cell is a seaweed cell. Preferred prokaryotic cells include those selected from the group consisting of a bacterium, a cyanobacterium, cells that contain a bacteriophage, and / or a virus. The nucleic acid sequence encoding the polypeptide of the microbial cell preferably contains a promoter that is functional in the host cell which optionally is an inducible promoter for example by broth components. Preferred microbial cells used in the methods are yeast cells, such as Saccharomyces cells. In another embodiment of the invention, a recombinant yeast cell that converts more than about 5% 20: 3 fatty acid substrate to a 20: 4 fatty acid product is provided. A fat comprising one or more polyunsaturated fatty acids is also provided. The amount of that one or more polyunsaturated fatty acids is about 0.3-30% of arachidonic acid, about 0.2-30% of dihomo-α-linolenic acid, and about 0.2-30% of α-linolenic acid. A preferred fat of the invention is one in which the ratio of arachidonic acid: dihomo-gamma-linolenic acid: gamalinolenic acid is from about 1.0: 19.0: 30 to 6.0: 1.0: 0.2. Another preferred embodiment of the invention is a pharmaceutical composition comprising the fats in a pharmaceutically acceptable carrier. In addition, a nutritional composition comprising the fats of the invention is provided. The nutritional compositions of the invention are preferably administered to a mammalian host parenterally or internally. A preferred composition of the invention for internal consumption is a formula for infants. In a preferred embodiment, the nutritional compositions of the invention are in liquid form or in solid form. The present invention also includes a method for desaturating a fatty acid, wherein the method comprises culturing a recombinant microbial cell of the invention under conditions suitable for the expression of a polypeptide encoded by the nucleic acid, wherein the host cell further comprises a substrate of fatty acid of the polypeptide. In a preferred embodiment, a desaturated fatty acid is provided by the method, including an oil comprising the fatty acid. The present invention is also directed to purified nucleotide and peptide sequences presented in SEQ ID NO: 1-34. The present invention is further directed to methods for using the sequences presented in SEQ ID NO: 1-34 as probes to identify related sequences, as components of expression systems and as components of systems useful for producing transgenic oil. The present invention is further directed to methods for obtaining biosystems of altered long-chain polyunsaturated fatty acids by cultivating transgenic microbes that encode transgene expression products that desaturate a fatty acid molecule at the carboxyl end carbon 5 of the acid molecule. fatty. The present invention is further directed to formulas, dietary supplements or dietary supplements in the form of a liquid or a solid containing the long chain fatty acids of the invention. These formulas and supplements can be administered to a human or an animal. The formulas and supplements of the invention may also comprise at least one macronutrient selected from the group consisting of coconut oil, soybean oil, canola oil, mono and diglycerides, glucose, edible lactose, electrodialyzed serum, electrodialyzed skim milk, milk, soy protein, and other protein hydrolysates. The formulas of the present invention may further include at least one vitamin selected from the group consisting of Vitamins A, C, D, E, and complex B; and at least one mineral selected from the group consisting of calcium, magnesium, zinc, manganese, sodium, potassium, phosphorus, copper, chlorine, iodine, selenium and iron. The present invention is further directed to a method of treating a patient having a problem caused by insufficient consumption or production of polyunsaturated fatty acids comprising administering to the patient a dietary substitute of the invention in an amount sufficient to effect the treatment of the patient. The present invention is further directed to cosmetic and pharmaceutical compositions of the material of the invention. The present invention is also directed to an isolated nucleotide sequence comprising a nucleotide sequence selected from the group consisting of: SEQ ID NO: 13 SEQ ID NO: 15; SEQ ID NO: 17; SEQ ID NO: 19; SEQ ID NO: 21 SEQ ID NO: 22; SEQ ID NO: 23; SEQ ID NO: 24; SEQ ID NO: 25 SEQ ID NO: 26 and SEQ ID NO: 27. The present invention also addresses a sequence of isolated peptides comprising a sequence of peptides selected from the group consisting of: SEQ ID NO: 14; SEQ ID NO: 16; SEQ ID NO: 18; SEQ ID NO: 20; SEQ ID NO: 28; SEQ ID NO: 29; SEQ ID NO: 30; SEQ -ID NO: 31; SEQ ID NO: 32; SEQ ID - NO: 33 and SEQ ID NO: 34. The present invention is further directed to transgenic oils in pharmaceutically acceptable carriers. The present invention is further directed to nutritional supplements, cosmetic substances and formulas for infants containing transgenic oils. The present invention is further directed to a method for obtaining biosynthesis of altered long-chain polyunsaturated fatty acids comprising the steps of: culturing a microbe that has cells containing a transgene encoding a transgene expression product which desaturates a fatty acid molecule at the carboxyl end carbon 5 of the fatty acid molecule, wherein the transgene is operably associated with an expression control sequence, under conditions by which the transgene is expressed, thereby altering the biosynthesis of long-chain polyunsaturated fatty acids in cells. The present invention is further directed to the use of polyunsaturated chain fatty acid from the group consisting of arachidonic acid, dihomo-gamma-linolenic acid and eicosapentaenoic acid. The present invention is also directed to pharmaceutical compositions comprising at least one nutrient selected from the group consisting of a vitamin, a mineral, a carbohydrate, a sugar, an amino acid, a free fatty acid, a phospholipid, an antioxidant, and a phenolic compound. Brief description of the Drawings Figure 1 shows the possible routes for the synthesis of arachidonic acid (20: 4? 5, 8, 11, 14) and stearidonic acid (18: 4? 6, 9, 12, 15) from palmitic acid (C16) from a variety of organisms, including algae, Mortierella and humans. These polyunsaturated fatty acids can serve as precursors to other molecules important to humans and other animals, including prostacyclines, leukotrienes, and prostaglandins, some of which are shown. Figure 2 shows possible routes for the production of polyunsaturated fatty acids in addition to arachidonic acid, including eicosapentaenoic acid and docosahexaenoic acid, for a variety of organisms. Figures 3A-D show the DNA sequence of the ? 5-desaturase from Mortierella alpina and the deduced amino acid sequence. Figure 4 shows the amino acid sequence of the polymerase chain reaction fragment (see Example 1). Figures 5A and 5B show alignments of the? 5-desaturase protein sequence with? 6-desaturases. Figures 6A and 6B show the effect of time of substrate addition relative to induction in the conversion of the substrate into product in SC334 containing the? 5-desaturase gene. Figures 7A and 7B show the effect of the inducer concentration on the expression of? 5-desaturase in SC334. Figures 8A and 8B show the effect of the induction temperature on the activity of? 5-desaturase in SC334. Figures 9A and 9B show the effect of the host strain on the conversion of substrate into product in strains expressing the? 5-desaturase gene at 15 ° C. Figures 10A and 10B show the effect-of the host strain on the conversion of substrate into product in strains expressing the gene 5-desaturase at 30 ° C. Figure 11 shows the effect of a host strain expressing choline transferase as well as the α 5-desaturase gene in the conversion of substrate into product. Figures 12A and 12B show the effect of media composition and temperature on the conversion of the substrate into product in two host strains expressing the? 5-desaturase gene. Figure 13 shows alignment of the protein sequence of Ma29 and contig 253538a. Figure 14 shows alignment of the protein sequence of Ma524 and contig 253538a.

Brief description of the sequence listings SEQ ID NO: 1 shows a DNA sequence of the? 5-desaturase from Mortierella alpina. SEQ ID NO: 2 shows an amino acid sequence of the? 5-desaturase from Mortierella alpina. SEQ ID NO: 3 shows the deduced amino acid sequence of the polymerase chain reaction fragment of M. alpina (See Example 1). SEQ ID NO: 4 - SEQ ID NO: 7 show the amino acid sequences deduced from several 6-desaturases. SEQ ID NO: 8 and SEQ ID NO: 9 show polymerase chain reaction primer sequences for α6-desaturases. SEQ ID NO: 10 shows a primer for reverse transcription of total RNA. SEQ ID NO: 11 and SEQ ID NO: 12 show amino acid motifs for desaturases sequences. SEQ ID NO: 13 and SEQ ID NO: 14 show the nucleotide and amino acid sequence for the desaturase sequence of Dictyostelium discoideum. SEQ ID NO: 15 and SEQ ID NO: 16 show the nucleotide and amino acid sequences of a desaturase sequence of P2-aeodacty2um tricornutum. SEQ ID NO: 17-20 show the nucleotide and deduced amino acid sequence of a Schizochytrium cDNA clone.

SEQ ID NO: 21-27 shows nucleotide sequences for human desaturases. SEQ ID NO: 28 - SEQ ID NO: 34 show peptide sequences for human desaturases. DETAILED DESCRIPTION OF THE INVENTION In order to ensure a complete understanding of the invention, the following definitions are provided: 5-Desaturase: 5-desaturase is an enzyme that introduces a double bond between the carbon atoms 5 and 6 of the carboxyl terminus of a fatty acid molecule. 6-Desaturase: 6-desaturase is an enzyme that introduces a double bond between the carbons 6 and 7 of the carboxyl end of a fatty acid molecule. ? 9-Desaturase:? 9-desaturase is an enzyme that introduces a double bond between carbons 9 and 10 of the carboxyl end of a fatty acid molecule. ? 12-Desaturase:? 12-desaturase is an enzyme that introduces a double bond between carbons 12 and 13 of the carboxyl end of a fatty acid molecule. Fatty Acids: Fatty acids are a class of compounds that contain a long hydrocarbon chain and a terminal carboxylate group. The fatty acids include the following: Taking these definitions into account, the present invention is directed to novel DNA sequences, DNA constructs, methods and compositions that are provided to allow the modification of the polyunsaturated long chain fatty acid content of, for example, microbial cells or animals. The host cells are engineered to express a sense or antisense transcript of a DNA encoding a polypeptide (s) that catalyzes the conversion of dihomo-gamma-linolenic acid to arachidonic acid. The substrate (s) for the expressed enzyme can be produced by the host cell or can be delivered exogenously. To achieve expression, the transformed DNA is operably associated with transcriptional and translational regulatory initiation and termination regions that are functional in the host cell. Constructs comprising the gene to be expressed can provide integration into the genome of the host cell or can replicate autonomously in the host cell. For the production of arachidonic acid, the generally used expression tapes include a tape that provides? 5-desaturase activity, particularly in a host cell that produces or can pick up dihomo-gamma-linolenic acid. The production of α6-unsaturated fatty acids, such as arachidonic acid, is favored in a host micro-organism or an animal that is substantially free of α-linolenic acid. The host is selected or obtained by removing or inhibiting the activity of a? 15 or? 3 desaturase (see Figure 2). The endogenous desaturase activity can be affected by providing an expression cassette for a? 15 or? 3 antisense transcript., by interrupting a target gene? 15 or? 3 through insertion, substitution and / or deletion of all or part of the target gene, or by adding a? 15 or? 3-desaturase inhibitor. The production of LA can also be increased by providing expression bands for? 9 and / or? L2-desaturases where their respective enzymatic activities are limiting. MICROBIAL PRODUCTION OF FATTY ACIDS The microbial production of fatty acids has several advantages over the purification of natural sources such as fish or plants. Many microbes are known with compositions of fats simplified to a large extent compared to those of higher organisms, simplifying the purification of the desired components. Microbial production is not subject to fluctuations caused by external variables such as climate and food supply. The fats produced microbially are substantially free of contamination by environmental contaminants. Additionally, microbes can provide polyunsaturated fatty acids in particular forms that may have specific uses. For example, Spirulina can provide predominantly polyunsaturated fatty acids in the first and third positions of triglycerides; digestion by pancreatic lipases preferentially releases fatty acids from these positions. After human or animal ingestion of triglycerides derived from Spirulina, these polyunsaturated fatty acids are released by pancreatic lipases as free fatty acids and are thus directly available, for example, for the development of the infant brain. Additionally, the production of microbial fats can be manipulated by controlling the conditions of the culture, especially by providing particular substrates for the microbially expressed enzymes, or by the addition of compounds that suppress undesired biochemical trajectories. In addition to these advantages, the production of fatty acids from recombinant microbes provides the ability to alter the profile of naturally occurring microbial fatty acids by providing new synthetic pathways in the host or by suppressing unwanted pathways, thus increasing the levels of fatty acids desired polyunsaturates, or conjugated forms thereof, and decreasing unwanted polyunsaturated fatty acid levels. PRODUCTION OF FATTY ACIDS IN ANIMALS The production of fatty acids in animals also has several advantages. The expression of desaturase genes in animals can produce greatly increased levels of polyunsaturated fatty acids in animal tissues, making recovery from these tissues more economical. For example, when desired polyunsaturated fatty acids are expressed in the breast milk of animals, methods for isolating polyunsaturated fatty acids from animal milk are well established. In addition to providing a source for the purification of desired polyunsaturated fatty acids, animal breast milk can be manipulated through the expression of desaturase genes, either alone or in combination with other human genes, to provide animal milks with a composition of polyunsaturated fatty acids substantially similar to human breast milk during the different stages of infant development. Humanized animal milks could serve as formulas for infants when it is impossible to breastfeed human or unwanted, or in cases of malnutrition or disease. Depending on the host cell, the availability of the substrate, and the desired end product, several polypeptides, particularly desaturases, are of interest. By "desaturase" is meant a polypeptide that can desaturate one or more fatty acids to produce mono or polyunsaturated fatty acid or precursor thereof of interest. Of particular interest are polypeptides that can catalyze the conversion of dihomo-gamma-linolenic acid to produce arachidonic acid which includes enzymes that desaturate at the 5-position. By "polypeptide" is meant any chain of amino acids, independent of the length or posttranslational modification, for example, glycosylation or phosphorylation. Considerations for choosing a specific polypeptide having desaturase activity includes the optimum pH of the polypeptide, whether the polypeptide is an enzyme that limits the regimen or a component thereof, or the desaturase used is essential for the synthesis of a poly fatty acid unsaturated unsaturated, and / or cofactors required by the polypeptide. The expressed polypeptide preferably has parameters compatible with the biochemical environment of its location in the host cell. For example, the polypeptide may have to compete for substrate with other enzymes in the host cell. Therefore, analyzes of the Km and the specific activity of the polypeptide in question are considered to determine the suitability of a given polypeptide for modifying the production of polyunsaturated fatty acids in a given host cell. The polypeptide used in a particular situation is one that can function under the conditions present in the intended host cell but can otherwise be any polypeptide having desaturase activity which has the desired characteristic of being able to modify the relative production of an acid desired poly-unsaturated fatty acid. For the production of arachidonic acid, the DNA sequence used encodes a polypeptide having? 5-desaturase activity. In particular cases, this can be coupled are an expression band that provides the production of a polypeptide that has β6-desaturase astivity and the host cell can optionally possess any α5-desaturase present, for example by providing a trans-siphonase for the production of antisense sesuensias to the transsripsion prodrug of the? 5-desaturase, by interrupting the? 5-desaturase gene, or using a host cell which naturally has, or has been mutated to have, low asthat? 15-desaturase. Inhibition of undesired desaturase pathways can also be brought to flavor through the use of specific desaturase inhibitors such as those described in U.S. Patent No. 4,778,630. The relaxation of the biosylation of syntheses used may depend in part on the profile of the polyunsaturated fatty acid of the host cell. When the astility? 5-desaturase of the host cell is limiting, overexpression of? 5-desaturase alone will generally be sufficient to provide the produssion of increased arachidonic acid increased in presensia of an appropriate substrate such as somo-dihomo-gama-linoleniso . The production of acid arachidonide can also be increased by providing expression syntaxes for? 9 or? 12-desaturase genes when the astivities of these desaturases are limiting. A schematic for the synthesis of the arachidid (20: 4? 5'8,11,14) from the arid palm (C16) is shown in Figure 1. A key enzyme in this route is the? 5-desaturase which is the bitter dihomo -? - linolenide (DGLA, acidic eicosatrienoiso) in the arid arachidonid. The sonification of α-α-linolenide (ALA) in solid stearidóniso by a β6-desaturase is also shown. The produssion of polyunsaturated fatty acids in addition to acidic arachidonide, including eicosapentaenoiso acid and acid dososahexaenoiso is shown in Figure 2. SOURCES OF POLIPEPTIDES THAT HAVE ACTIVITY DESATURASE A source of polypeptides having astatura desaturase and oligonusleotides that sodify these polypeptides are organisms that produce a desired poly-unsaturated fatty acid. As an example, microorganisms that have a capacity to produce arachidonic acid can be used as a source of asthma? 5-desaturase. These misro-organisms include, for example, those belonging to the genera Mortierella, Conidiobolus, Pythium, Phytophathora. Penisillium, Porphyridium, Coidosporium, Musor, Fusarium, Aspergillus, Rhodotorula, and Entomophthora. Where the Porphyridium genus, of particular interest is Porphyridium sruentum. Within the Mortierella genus, Mortierella elongata, Mortierella exigua, Mortierella hyarophila, Mortierella ramanniana, var. angulispora, and Mortierella alpina.

Within the Mucor genus, of particular interest are Mucor circinelloides and Mucor javanicus. The DNAs coding for desired desaturases can be identified in several ways. As an example, a desired desaturase source, for example genomic libraries or cDNA libraries for Mortierella, are selected are enzymatically or chemically detectable probes synthesized, which can be made from DNA, RNA, or nucleotides that do not occur naturally or mixtures of the same. The probes can be synthesized systematically from DNA of desaturases known for normal or reduced stringency methods. Oligonucleotide probes can also be used to analyze sources and can be based on sequences of known desaturases, including the sequencies conserved between desaturases conosidas, or in sesuencias of peptides contained from the desired purified protein. Oligonucleotide probes based on amino acid sequences can degenerate to encompass the degeneracy of the genetic code, or they can tilt in favor of the preferred codons of the source organism. Oligonucleotides can also be used as primers for polymerase chain reaction from reverse transcribed mRNA from a known or presumed source; the polymerase chain reaction product can be full-length cDNA or can be used to generate a probe to obtain the desired full-length cDNA. Alternatively, a desired protein can be severely sessed and the total synthesis of the DNA that sodifices that polypeptide can be brought to taste. In the meantime, the desired DNA or genome has been isolated, it can be sessed by sonoside methods. It is resumed in the case that these methods are subject to errors, such as multiple sesuensiamiento of the same region somo routine and is still expected to sondusir to measurable regimes of errors in the resulting dedusida sesuensia, particularly in regions that have repeated domains, estrustura sesundaria extensive, or unusual base sompositions, such somo regions are high sustained GC base. When dissrepansias arise, ressuensiamiento can be made and special methods can be used. Species methods can include altering the conditions of sequencing using: different temperatures; different enzymes; proteins that alter the sapsity of the oligonucleotides to form higher-order estrusturas; altered nusleotides such as ITP or methylated dGTP; different gel sompositions, for example formamide added; different selectors or selectors, located at different distances from the problem region; or different patterns such as DNA from a single chain. The mRNA sesuensiamiento can also be used. For the most part, some or all of the sodifisation sesuensia for the polypeptide having astatura desaturase is from a natural source. In some situations, however, it is desirable to modify all or a portion of the codons, for example, to increase expression, using the preferred codons of hosts. Preferred guest hosts can be determined from the freshest serums in the proteins expressed in the largest holiness in a particular host species of interest. In this way, the coding sequence for a polypeptide having astatura desaturase can be synthesized in whole or in part. All or DNA splices can also be synthesized to remove any destabilizing sesuensia or regions of secondary strustence that would be present in the transsomal mRNA. All or portions of the DNA can also be synthesized to alter the composition of the base to one or more preferable in the desired host cell. The methods to synthesize sesuensias and put the sesuensias together are very well established in the literature. Mutagenesis and senesion in vitro, site-directed mutagenesis, or other elements can be used to obtain mutations of naturally occurring desaturase genes to produce a polypeptide having desaturase activity in vivo with physical parameters and more desirable kinetics to function in the host cell, such as a longer half-life or a regimen greater of produssion of a desired poly-unsaturated fatty acid. Mortierella alpina desaturase Of particular interest is the 5-desaturase of Mortierella alpina which has 446 amino acids; the amino acid sequence is shown in Figure 3. The gene encoding the 5-desaturase of Mortierella alpina can be expressed in micro-organisms or transgenic animals to effect greater synthesis of arachidonic acid from dihomo-gamma-dihydrate. linolenic Other DNAs that are substantially identical to the? 5-desaturase DNA of Mortierella alpina, or that encode polypeptide that are substantially identical to the? 5-desaturase polypeptide of Mortierella alpina can also be used. By "substantially identical" is meant an amino acid sequence or a nucleic acid sequence which exhibits in order of increasing preference at least 60%, 80%, 90% or 95% homology to the amino acid sequence or acid sequence 5? desaturase nucleic acid from Mortierella alpina encoding the amino acid sequence. For polypeptides, the length of the comparison sequences is at least 16 amino acids, preferably at least 20 amino acids, or more preferably 35 amino acids. For nucleic acid, the length of comparison sequences is generally at least 50 nucleotides, preferably at least 60 nucleotides, and more preferably at least 75 nucleotides, and more preferably 110 nucleotides. Homology is typically measured using sequence analysis software, for example the sequence analysis software package from the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wisconsin 53705, MEGAling (DNAStar, Inc., 1228 S. Park St., Madison, Wisconsin 53715), and MacVector (Oxford Molecular Group, 2105 S. Bascom Avenue, Suite 200, Campbell, California 95008). This software compares similar sequences by assigning degrees of homology to various substitutions, deletions, and other modifications. Conservative substitutions typically include substitutions within the following groups: glycine and alanine; valine, isoleucine and leucine; aspartic acid, glutamic acid, asparagine, and glutamine; serine and threonine; lysine and arginine; and phenylalanine and tyrosine. Substitutions may also be based on the hydrophobicity or hydrophilicity conserved (Kyte and Doolittle, J. Mol. Biol. 157: 105-132, 1982), or on the basis of the ability to assume secondary structure of similar polypeptide (Chou and Fas an, Adv * Enzymol 47: 45-148, 1978). Other desaturases Included in the present invention are the related desaturases thereof or other organisms. These related desaturases include variants of the described? 5-desaturase that occur naturally within the same or different Mortierella species as well as homologs of the? 5-desat.urase described from other species. Also included are desaturases which, while not substantially identical to the 5-desaturase of Mortierella alpina, desaturate a fatty acid molecule at the carbon 5 of the carboxyl end of the fatty acid molecule. The related desaturases can be identified by their ability to function substantially the same as the described desaturases; that is, they are still able to effectively convert dihomo-gamma-linolenic acid into arachidonic acid. Related desaturases can also be identified by analyzing sequence databases to determine sequences homologous to the desaturase described, by hybridizing a probe based on the described desaturase to a library constructed from the "source" organism, or by RT-PCR using mRNA from the source organism and primers based on the desaturase described.These desaturases include those of humans, Dictyostelium discoideum and Phaeodactylum tricornum.The regions of a desaturase polypeptide important for the desaturase activity can be determined through routine mutagenesis, expression of the Resulting mutant polypeptides and determination of their activities Mutants may include deletions, insertions and specific mutations, or a combination thereof. A typical functional analysis begins with deletion mutagenesis to determine the N and C terminal limits of the protein necessary for the function, and then internal deletions, insertions or point mutants are determined to further determine the regions necessary for the funtion. Other techniques can also be used, such as synthesis mutagenesis or total synthesis. Deletion mutagenesis is carried out by, for example, using exsnusleases to sequentially remove the coding regions 5 f or 3 l. There are games available for these teens. After suppression, the sodifisation region is sompleta ligand oligonusleotides that are either inisium or detensioning sodons to the deleted sodifisation region after suppression 57 or 3, respectively. Alternatively, the oligonucleotides that sodiate inisium or detensioning sodons are inserted into the sodifisation region by a variety of methods that include site-directed mutagenesis, mutagenized polymerase chain reaction or by ligase on the digested DNA at restriction sites existing Internal deletions can be made similarly through a variety of methods that include the use of existing restriction sites in the DNA, through the use of mutagenized primers via site-directed mutagenesis or mutagenicity polymerase saase reassignment. The insertions are made through methods such as linker mutagenesis, site-directed mutagenesis, or mutagenase polymerase saase reassay. Point mutations can be achieved through techniques such as site-directed mutagenesis or mutagenicity polymerase saase reassessment. Mutant mutagenesis can also be used to identify regions of a desaturase polypeptide important for activity. A mutated construstion is expressed, and the sapsity of the resulting altered protein is tested to func- tion as a desaturase. This structure-function analysis can determine which regions can be replaced, their regions tolerate insertions, and which point mutations allow the mutant protein to function sustantially in the same way as the original desaturase. All these mutant proteins and sequences of nusleotides that sodifisan are within the alsanse of the present invention. EXPRESSION OF DESATURASE GENES The DNA that sodifies a desaturase polypeptide, solves in a sapphine dressing of a refill in a host cell, or propagates in vitro by means of tsetse such as polymerase saphenous reassessment or saphenous reassessment has been obtained. of long polymerase. The repellant vesicles can include plasmids, phages, viruses, semites and the like. Desirable markers include those useful for mutagenesis of the gene of interest or for the expression of the gene of interest in the host cells. The long-chain polymerase chain reaction technique has made possible the in vitro propagation of large sequences, so that modifications of the gene of interest, such as mutagenesis or addition of the expression signals, and the propagation of the resulting constructs are possible. it can present entirely in vi tro without the use of a replicating vector or in a host cell. For the expression of a desaturase polypeptide, the transcription and functional translation initiation and termination regions are operably linked to the DNA encoding the desaturase polypeptide. The expression of the polypeptide coding region can be carried out in vi tro or in a host cell. The transcription and translation initiation and termination regions are derived from a variety of non-exlusive sources, including DNA to be expressed, genes known or suspected to be capable of expression in the desired system, expression vectors, chemical synthesis, or from an endogenous site in a host cell. Expression In Vitro Expression in vi tro can be carried out, for example, by singling out the sodifisation region for the desaturase polypeptide in an expression vector designated for in vitro use and adding used reticulocite and cofactors; Labeled amino acids can be incorporated if desired. These in vitro expression vectors can provide some or all of the expression signals needed in the system used. These methods are well known in the art and the components of the system are commercially available. The reaction mixture can be tested directly for the polypeptide, for example by determining its activity, or the synthesized polypeptide can be purified and then tested. Expression in a host cell Expression in a host cell can be carried out in a transient or stable manner. Transient expression can be presented from introduced construssions which contain functional expression signals in the host cell, but construssions do not re-aggregate and rarely integrate into the host cell, or where the host cell does not proliferate. Transient expression can also be carried out by inducing the activity of a regulatable promoter operably linked to the gene of interest, although these inducible systems often exhibit a low basal level of expression. Stable expression can be achieved by introducing a construct that can be integrated into the host genome or that replicates autonomously in the host cell. Stable expression of the gene of interest can be selected through the use of a selectable marker located or transfected with the expression construct, followed by the selection of cells expressing the marker. When the expression is a result of integration, the integration of constructs can be randomly presented within the host genome or can be directed through the use of constructs containing regions of homology with the host genome sufficient to direct recombination with the host site. When the constructs are directed to an endogenous site, the endogenous site can provide all or some of the regulatory transcription and translation regions. When increased expression of the polypeptide desaturase in the source organism is desired, several methods can be employed. Additional genes that sodify the desaturase polypeptide can be injected into the host organism. The expression of the original desaturase site can also be increased through homologous recombination, for example by inserting a stronger promoter into the host genome to cause increased expression, by removing destabilizing sequences from either the mRNA or the encoded protein by deleting that information of the host genome, or by adding stabilizing sequences to the mRNA (U.S. Patent Number: 4,910,141). When it is desired to express more than one different gene, suitable regulatory regions and methods of expression, genes introduced into the host cell can be propagated through the use of replicating vectors or through integration into the host genome. When expressing two or more genes of separate replication vectors, it is desirable that each vector have a different replication medium. Each construction introduced, whether integrated or not, must have a different means of selection and must have no homology with the other constructions to maintain stable expression and avoid the reclassification of elements between constructions. Judicious choices of regulatory regions, selection means and propagation methods of the introduced construssion can be determined experimentally so that all the introduced genes are expressed at the levels necessary to provide the synthesis of the desired products. As an example, wherein the host cell is a yeast, transsripsion and funsional tradussion regions are provided in the yeast cells, particularly from host species. The regulatory regions of transrision initiation can be obtained, for example from genes in the glisolitis tray, such as alcohol dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase (GPD), phosphoglucoisomerase, phosphoglycerate kinase, etc. or regulatable genes such as acid phosphatase, lactase, metallothionein, glucoamylase, etc. Any of several regulatory sequences can be used in a particular situation, depending on whether constitutive or induced transcription is desired, the particular efficiency of the promoter in conjunction with the open reading frame of interest, the ability to bind a strong promoter with a region of control from a different promoter which allows for inducible transcription, ease of construsion, and the like. Of particular interest are the promoters that are astivan in the galastosa presensia. The inducible galactose promoters (GAL1, GAL7, and GALIO) have been used extensively for high-level expression and regulated protein expression in yeast (Lue et al., Mol.Cell. Biol. Vol. 7, p.3446, 1987; Johnston, Microbiol. Rev. Vol. 51, p. 458, 1987). The GAL promoter transcription is activated by the GAL4 protein, which binds to the promoter region and activates transcription when galactose is present. In the absence of galactose, the GAL80 antagonist binds to GAL4 and prevents GAL4 from activating transcription. The addition of galactose prevents GAL80 from inhibiting activation by GAL4. The nucleotide sequences surrounding the ATG translation initiation codon have been found to affect expression in yeast cells. If the desired polypeptide is poorly expressed in the yeast, the nucleotide sequences of exogenous genes can be modified to include an efficient yeast translation initiation sequence to obtain optimal expression of the gene. For expression in Saccharomyces, this can be done by site-directed mutagenesis of a gene expressed inefficiently by fusing it within the frame to an endogenous Saccharomyces gene, preferably a highly expressed gene, such as a lactase gene. The termination region can be derived from the 3 'region of the gene from which the initiation region is obtained or from a different gene. A large number of termination regions are known and have been found to be satisfactory in a variety of hosts of the same and different genera and species. The termination region is usually selected more as a matter of convenience rather than due to some particular property. Preferably, the termination region is derived from a yeast gene, particularly Saccharomyces, Schizosaccharomyces, Candida or Kluyveromyces. The 3 'regions of two mammalian genes, and interferon and QII2 interferon, are also known to work in yeast. INTRODUCTION OF BUILDINGS IN HOSPITAL CELLS Constructs that comprise the gene of interest can be introduced into a host cell by standard techniques. These techniques include transformation, protoplast fusion, lipofection, transfection, transduction, conjugation, infection, ballistic impact, electroporation, micro-injection, scraping, or any other method that introduced the gene of interest in the host cell. Transformation methods that are used include transformation by lithium acetate (Methods in Enzymology, Vol 194, p 186-187, 1991).

Conveniently, a host cell that has been manipulated by any method to pick up a DNA sequence or construct will be called in the present "transformed" or "recombinant". The subject host will have at least one copy of the expression construct and can have two or more, depending on whether the gene is integrated into the genome, amplified, or is present in an extrachromosomal element that has multiple copy numbers. When the host is a yeast, four major types of plasmid vectors can be used: yeast integration plasmids (YIps), j plasmids and yeast replication (YRps), yeast centromere plasmids (YCps), and yeast episomal plasmids (YEps). The yeast integrating plasmids lack a yeast replication origin and must be propagated as integrated elements in the yeast genome. Yeast replication plasmids have a replication sequence that is autonomously chromosomally derived and propagate as unstable, autonomously replicating plasmids of intermediate copy number (20 to 40). Plasmids from yeast centromeres have both a replication origin and a sentromer sesuensia and propagate as stably secreting, autonomously replicating plasmids of low copy number (10-20). The yeast episomal plasmids have a replication origin of the yeast 2μm plasmid and propagate as irregular, autonomously replicating, high copy number plasmids. The presence of the plasmids in the yeast can be ensured by maintaining the selection of a marker in the plasmid. Of particular interest are the yeast vectors pYES2 (a YEp plasmid available from Invitrogen, conferring uracil prototrophy and an GAL1 galactose inducible promoter for expression) pRS425-pGl (a YEp plasmid obtained from Dr. TH Chang, assistant professor of Molecular Genetics , Ohio State University, which contains a constitutive GPE promoter and confers leucine prototrophy), and pYX424 (a YEp plasmid that has a constitutive TP1 promoter conferring leucine prototrophy; Alber, T. and Kawasaki, G. (1982). " Mol. &Appl. Genetics 1: 419) .The transformed host cell can be identified by selection for a marker synthesized in the introduced construct.Alternatively, a marker construct can be introduced separately with the desired construct since many techniques Many transformed DNA molecules are introduced into host cells, typically transformed hosts are selected for their ability to to pray in selective media. The selective media can incorporate an antibiotic or lack a necessary factor for the growth of the non-transformed host, such as a nutrient or desorption factor. A marker gene introduced therefor can confer antibiotic resistance, or encode an essential growth factor or enzyme, and allow growth in selective media when expressed in the transformed host. The selection of a transformed host can also occur when the expressed marker protein can be detected, either directly or indirectly. The marker protein can be expressed alone or as a fusion to another protein. The marker protein can also be detected by its enzymatic activity; for example ß galactosidase can convert the X-gal substrate into a colored product, and the luciferase can convert the lusiferin into a product that emits light. The marker protein can be detected by its characteristics of producing light or modifying light; For example, the green fluorescent protein of Aeguorea victoria fluoresce suando lights are blue light. Antibodies can be used to detect the marker protein or a molecular tag in, for example, a protein of interest. Cells that express the marker protein or label can be selected, for example, visually, or by techniques such as FACS or washing using antibodies. For the selection of yeast transformants, any marker that works in yeast can be used. Desirably, resistance to kanamycin and aminoglycoside G418 are of interest, as well as the ability to grow in media lacking uracil, leucine, lysine or tryptophan. The production of? 5-desaturase of polyunsaturated fatty acids can be carried out either in prosariotis or eusariotis host cells. The prosariottic cells of interest include Essherrshia, Basillusf Lastobasillus, Cyanobasteria and the like. Eukaryotic cells include mammalian cells such as those of lactating animals, bird cells such as chickens, and other cells amenable to genetic manipulation including insect cells, fungi, and algae. The cells can be cultured or formed as part or all of a host organism including an animal. Viruses and bacteriophages can also be used with cells in the production of polyunsaturated fatty acids, particularly for gene transferensia, selction digestion and senession. In a preferred embodiment, the host is any micro-organism or animal that produces dihomo-gamma-linolenic acid and / or can assimilate exogenously supplied dihomo-gamma-linolenide acid, and preferably produces great sanities of dihomo-gamma-linolenide. Examples of host animals include mice, rats, sonejos, chickens, sodornises, turkeys, bovines, sheep, serdos, sabras, yaks, ets. , which are detrimental to genetic manipulation and sloning for the rapid expansion of the population expressed by the transgene. For animals, a transgene 5-desaturase can be adapted for its expression in target organelles, tissues and sorporal fluids through the modification of the regulatory regions of the gene. Of particular interest is the production of polyunsaturated fatty acids in the breast milk of the host animal. Expression in yeast Examples of host micro-organisms include Saccharomyces cerevisiae, Saccharomyces carlsbergensis, or other yeasts such as Candida, Kluyveromyces or other fungi such as for example filamentous fungi such as Aspergillus somo, Neurospora, Penicillium, ets. The desirable characteristics of a host microorganism are, for example, that it is genetically well characterized, that it can be used for the high level expression of the product using ultra high density fermentation, and it is on the list of generally safe resounds as well (GRCS ) since the proposed final product is intended for ingestion by humans. Of particular interest is the use of a yeast, more particularly baker's yeast (S. cerevisae), as a host cell in the present invention. Strains of particular interest are SC334 (Mat. Pep4-3 prbl-1122 ura3-52 leu2-3,112 regl-501 gall, Gene 83: 57-64, 1989, Hovland P. and collaborators), YTC34 (a ade2-101 his3? 200 lys2-801 ura3-52; obtained from Dr. TH Chang, Ass Professor of Molecular Genetics, Ohio State University), YTC41 (a / o; ura3-52 / ura3 = 52 Iys2-801 / lys2-801 ade2-101 / ade2-101 trpl-? l / trpl-? l his3? 200 / his3? 200 leu2? l / leu2? l; obtained from Dr.

T. H. Chang, Ass. Professor of Molecular Genetics, Ohio State University), BJ1995 (obtained from the Yeast Genetic Stock Center, 1021 Donner Laboratory, Berkeley, CA 94720), INVSC1 (Mat a his3? L leu2 trpl-289 ura3-52, obtained from Invitrogen, 1600 Faraday Ave., Carlsbad, CA 92008) and INVSC2 (Mat a his 3? 200 ura3-167; obtained from Invitrogen). Expression in bird species To produce polyunsaturated fatty acids in bird species and cells, such as chickens, turkeys, quails and ducks, gene transfer can be carried out by introducing a sequence of nucleic acids encoding? 5-desaturase in the cells following procedures known in the art. If a transgenic animal is desired, pluripotent stem cells of embryos can be provided with a vector carrying a? -desaturase encoding the transgene and developing in an adult animal (US Pat. No. 5,162,215; Ono et al. (1996) Comparative Biochemistry and Physiology A 133 (3): 287-292; WO 9612793; WO 9606160). In most cases, the transgene will be modified to express high levels of the desaturase in order to increase the production of polyunsaturated fatty acids. The transgene can be modified, for example, by providing transcriptional and / or translational regulatory regions that function in avian cells, such as promoters that direct expression in particular tissues and parts of the egg such as the yolk. The regulatory regions of the gene can be obtained from a variety of sources, including chicken anemia or bird leukosis viruses or bird genes such as chicken ovalbumin gene. Expression in insect cells The production of polyunsaturated fatty acids in insect cells can be carried out using baculovirus expression vectors carrying a? 5-desaturase transgene.

Baculovirus expression vectors are available from various commercial sources such as Clonetech. Methods for producing hybrid and transgenic strains of algae, such as seaweed, which contain and express a desaturase transgene, are also provided. For example, transgenic marine algae can be prepared as disclosed in U.S. Patent Number: 5,426,040. With or with other expression systems described above, the timing, extent of expression and activity of the desaturase transgene can be regulated by adjusting the sequencing of polypeptide coding is the appropriate transcriptional and translational regulatory regions selected for a particular use. Of particular interest are the promoter regions that can be induced under previously selected growth conditions. For example, the introduction of mutations responsive to temperature sensitivity and / or metabolites in the coding sequences of the desaturase transgene, its regulatory regions, and / or the genome of the cells into which the transgene is introduced can be used for this purpose Expression in plants The production of polyunsaturated fatty acids in plants can be carried out using various plant transformation systems such as the use of Agrobacterium tumefaciens, plant viruses, transformation of cells into particles and the like which are disclosed in the applicant's reissued applications Patent applications of the United States of America with serial numbers 08 / 834,033 and 08 / 956,985 and partial continuation requests filed simultaneously with this application which is incorporated herein by reference. The transformed host cell is cultured under suitable conditions adapted for a desired final result. In order for the host cells to grow in culture, the conditions are typically optimized to produce the higher and more economical yield of polyunsaturated fatty acids, which are related to the selected desaturase activity. The conditions of the media that can be optimized include: carbon source, nitrogen source, substrate addition, final concentration of the added substrate, added substrate form, aerobic or anaerobic growth, culture temperature, inducing substance, induction temperature, phase of srecimiento in the induction, phase of growth in harvest, pH, density, and maintenance of selection. Mere-organisms such as yeasts, for example, are preferably cultured using selected media of interest, including yeast peptone broth (YPD) and minimal media (which are amino acids, yeast nitrogen base, and ammonium sulfate). , and lacks a component by selection, for example uracil). Desirably, the substrates to be added are first dissolved in ethanol. When necessary, expression of the polypeptide of interest can be induced, for example by including or adding galactose to induce the expression of a GAL promoter. Expression in an animal The expression of cells of a host animal can also be carried out in a transient or stable manner. Transient expression can be carried out via known methods, for example infection or lipofection, and can be repeated in order to maintain desired expression levels of the introduced construct (see Ebert, TCP publication WO 94/05782). Stable expression can be carried out via integration of a construssion into the host genome, resulting in a transgenic animal. Construstion can be introduced, for example, by micro-injection of the construction into the pronucleus of a fertilized egg, or by transfection, retroviral infection or other techniques whereby the construction is introduced in a line-sealing that can be formed or insorporate in an adult animal (Patent of the United States of North America number 4,873,191; Patent of the United States of North America number 5,530,177; Patent of the United States of North America number 5,565,362; Patent of the United States of North America number 5,366,894; Wilmut and collaborators (1977) Nature 385: 810). Respecting eggs or embryos are transferred to a surrogate mother (Patent of the States United States of North America number 4,873,191; Patent of the United States of North America number 5,530,177; Patent of the United States of North America number 5,366,894; Wilmut and collaborators (supra)). After hatching, the transgenic animals are identified, for example, by the presence of an introduced marser gene, such as the skin solor, or by re-sorting of polymerase chain or Southern tinsion from a blood sample, leshe or tissue to detest the introduction of blood, or by an immunological or enzymatic assay to detest the protein expressed by the products produced therefrom (U.S. Patent No. 4,873,191; U.S. Patent No. 5,530,177 U.S. Patent No. 5,565,362; U.S. Patent No. 5,366,894; "Wilmut et allaborators" (supra)); The resulting transgenic animals can be completely transgenic or can be mosaisos, having the transgenes in only a subset of their cells. The arrival of mammalian slonasión, taken to flavor by fusion of a nusleada cell is an egg not nusled, followed by transferensia in a surrogate mother, presents the possibility of rapid produssión, to great essala, after obtaining a "founder" animal or cell that presents the introduced strutssion; before this, it was necessary for the transgene to be present in the germline of the animal for its propagation (Wilmut and co-workers (supra)). The expression in a host animal presents / displays sieves efisiensias, partisularly suando the guest is an animal domestisado. For the production of polyunsaturated fatty acids in a fluid readily obtainable from the host animal, such somo leshe, the desaturase transgene can be expressed in mammary cells from a female host, and alter the saturation of polyunsaturated fatty acid of the host cells. The desaturase transgene can be adapted for expression so that it is retained in the mammary cells, or is sesreted in the leshe, to form the produtos of the reassumption of polyunsaturated fatty acids located in the milk (PCT publisasión WO 95/24488 ). Expression can be targeted for expression in mammary tissue using thiosyllabic regulatory sesuensias, such as a-lastalbumin, α-casein, β-sasein, β-seasein, β-seasein, β-bovine lovastatin, or whey protein. milk, and optionally may include one or more introns and / or secretory signal sequences (U.S. Patent No. 5,530,177; Rosen, U.S. Patent No. 5,565,362; Clark et al., U.S. Pat. North America number 5,366,894; Garner et al., PCT publication WO 95/23868). The expression of transgenes desaturase, or antisense desaturase transcripts, adapted in this manner can be used to alter the levels of fatty polyunsaturated fatty acids, or derivatives thereof, embedded in the leshe of the animals. Additionally, the transgene? -desaturase can be expressed either on its own or are other transgenes, are for the purpose of producing animal leshe containing higher propsions of desired polyunsaturated fatty acids or proportions of polyunsaturated fatty acids and concentrations that resemble human breast milk (Prieto et al., PCT publication WO 95/24494). PURIFICATION OF FATTY ACIDS The desaturated fatty acids in position? 5 can be found in the micro-organism or host animal as free fatty acids or in conjugated forms such as acylglycerols, phospholipids, sulpholipids or glycolipids, and can be extracted from the host cell through a variety of means well known in the art. These means may include extraction with organic solvents, sonication, extraction of supercritical fluid using for example carbon dioxide, and physical means such as presses, or combinations thereof. Of particular interest is the extraction with methanol and chloroform. When desired, the aqueous layer can be acidified to protonate negatively charged fractions and thereby increase the partition of the desired products in the organic layer. After extraction, the organic solvents can be removed by evaporation under a stream of nitrogen. When isolated in conjugated forms, the products can be enzymatically or chemically dissociated to release the free fatty acid or a less complex conjugate of interest, and then can be subjected to other manipulations to produce a desired final product. Desirably conjugated forms of fatty acids are dissociated with potassium hydroxide. If further purification is necessary, standard methods can be employed. These methods may include extraction, urea treatment, fractional crystallization, high performance liquid chromatography, fractional distillation, silicon gel chromatography, centrifugation or high speed distillation, or combinations of these techniques. The protection of reactive groups, such as acid groups or alkenyls, can be done at any step by known techniques, for example alkylation or iodination. The methods used include methylation of the fatty acids to produce methyl esters. Similarly, protection groups can be removed at any step. Desirably, the purification of fractions containing arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid can be carried out by treatment with urea and / or fractional distillation. USES OF FATTY ACIDS There are several uses for the fatty acids of the present invention. DNA-based probes of the present invention can be found in methods for isolating related molecules or in methods for detecting organisms that express desaturases. When used as probes, the DNAs or oligonucleotides must be detectable. This is usually carried out by attaching a label to either an internal site, for example via the incorporation of a modified residue, or in the 5 'or 3' term. These labels can be directly detectable, can be attached to a secondary molecule that is detectably labeled, or can be linked to an unlabeled secondary molecule and a detectably labeled terrestrial molecule; this process can be extended as long as it is practical to achieve a satisfactorily detectable signal without unacceptable levels of background signal. Secondary, tertiary or bridging systems may include the use of antibodies directed against any other molecule, - including labels or other antibodies, or may involve molecules that are linked together, for example a biotin-streptavidin / avidin system. Detectable labels typically include radioactive isotopes, molecules that produce chemically or enzymatically or alter light, enzymes that produce detectable reaction products, magnetic molecules, fluorescent molecules or molecules whose fluorescence or light emitting characteristics change after binding. Examples of labeling methods can be found in the United States Patent Number: 5,011,770. Alternatively, the binding of target molecules can be detected directly by measuring the change in heat of the solution at the junction of one. probe to the target via isothermal titration calorimetry, or by coating the probe or target on a surface and detecting the change in light scattering of the surface produced by attaching a target or probe, respectively, as can be done with the BIAcore system. The polyunsaturated fatty acids produced by recombinant elements find applications in a wide variety of areas. The supplementation of humans or animals with polyunsaturated fatty acids in various forms can result in increasing levels not only in polyunsaturated fatty acids, but also in their metabolic progeny. For example, when the inherent? -desaturase pathway does not work in an individual, treatment with arachidonic acid may result not only in increasing the levels of arachidonic acid, but also in products downstream from arachidonic acid such as prostaglandins (see Figure 1). Complex regulatory mechanisms may make it desirable to combine several polyunsaturated fatty acids, or add different conjugates of polyunsaturated fatty acids, in order to avoid, control or overcome these mechanisms to achieve the desired levels of specific polyunsaturated fatty acids. in an individual. NUTRIMENTAL COMPOSITIONS The present invention also includes nutritional compositions. These compositions, for purposes of the present invention, include any food or preparation for human consumption that is included for enteral or parenteral consumption, which when taken in the body (a) serves to nourish or build tissues or supply energy and / or (b) maintain, restore or support adequate nutritional status or metabolic function. The nutritional composition of the present invention comprises at least one fat or acid produced in accordance with the present invention and may be in either solid or liquid form. Additionally, the composition may include edible macronutrients, vitamins and minerals in desired amounts for a particular use. The amount of these ingredients will vary depending on whether the somposisance is intended for use are normal, healthy, children or adults who have espesialized needs such as those that accompany metaboleal metabolisms (for example, metabolic disorders). Examples of the macronutrients that can be added to the somposision include, but are not limited to, somatic fats, sarbohydrates, and proteins. Examples of these somatic fats include, but are not limited to, coso sauces, soybean oil, and mono and diglycerides. Examples of these sarbohydrates include, but are not limited to, glusosa, somestible cheese and hydrolyzed corn starch. Additionally, examples of proteins that can be used in the nutrimental somposision of the invention include but are not limited to soy proteins, electrodialyzed whey, electrodialyzed skim milk, buttermilk, or hydrolysates of these proteins. With respect to vitamins and minerals, the following can be added to the nutritional conditions of the present invention: salsium, phosphorus, potassium, sodium, sloro, magnesium, manganese, iron, on, zins, selenium, iodine, and vitamins A, E, D, C and the B-complex. Other vitamins and minerals can also be added. The somponents used in the nutrimental sompositions of the present invention will be of semi-purified or purified origin. By semi-purified or purified is meant a material that has been prepared by purification of a natural material or by synthesis. Examples of nutritional compositions of the present invention include but are not limited to infant formulas, dietary supplements, and rehydration compositions. Nutrient compositions of particular interest include but are not limited to those used for enteral or parenteral supplementation for infants, specialized formulas in infants, supplements for geriatrics, and supplements for those with gastrointestinal difficulties and / or malabsorption. Nutrient Compositions A typical nutritional composition of the present invention will contain edible macronutrients, vitamins and minerals in desired amounts for a particular use. The amounts of these ingredients will vary depending on whether the formulation is intended for use with normal, healthy individuals, temporarily exposed to stress, or subjects having specialized needs due to certain chronic or acute disease states (e.g., metabolic disorders). ). A person skilled in the art will understand that the components used in a nutritional formulation of the present invention are of semi-purified or purified origin. By semipurified or purified is meant a material that has been prepared by purification of a natural material or by synthesis. These tisnishes are very prominent in teasy (see, for example, Code of Federal Regulations for Food Ingredients and Food Prosessing; Resommended Dietary Allowanses (Code of Federal Regulations for Food Ingredients and Food Processing, recommended dietary sanctities), dth edition, National Academy Press, Washington, D.C., 1989). In a preferred embodiment, a nutritional composition of the present invention is an enteral nutritional prodrug, more preferably an enteral nutritional prodrug for an adult or child. Of sonformity they are the previous thing in another aspesto of the invention, it is provided a nutritional formula that is suitable for feeding adults, who are experiencing stress. The formula includes, in addition to the polyunsaturated fatty acids of the invention, macronutrients, vitamins and minerals in sanctities designed to provide the daily nutritional requirements of adults. The somnutrient somatic elements include somatic fats, sarbohydrates and proteins. Exemplary somatic fats include bland, soy, mono and diglycerides and polyunsaturated fatty foods of this invention. Examples of sarbohydrates are glusosa, somestible pliant and hydrolyzed corn starch. A typical protein source would be soy protein, electrodialyzed serum or lesion dessremada elestrodializada or serum of leshe, by the hydrolysates of these proteins, although other sources of proteins are also available and can be used. These macronutrients would be added in the form of nutrient compounds commonly asepted in sanctity equivalent to those of the human leshe or an energy base, it is desir, they are base by salor. The methods for liquid and enteral nutrient formulas are very sonoside in the teasant and they are dessriben in detail in the examples. The enteral formula can be sterilized and subsequently used in a ready-to-feed or soured base in a liquid or powder. The powder can be prepared by drying the enteral formula prepared as indicated above, and the formula can be reconstituted by rehydrating the sonsentrate. Nutrient formulas for adults and lactants are very useful in the tea and are available (for example, Similas®, Ensure®, Jevity® and Alimentum® from Ross Produsts Division, Abbott Laboratories). One example or one of the present invention can be added to any of these formulas in the sanctities described below. The energy density of the nutrient supersaturation is in liquid form, typically ranging from about 0.6 kilocalories to 3.0 kilocalories per milliliter.When in solid or powder form, the nutritional supplement can contain from about 1.2 to over 9. kilocalories per gram, preferably 3 to 7 kilocalories per gram In general, the osmolality of a liquid product should be less than 700 mOsm and more preferably less than 660 mOsm The nutritional formula would typically include vitamins and minerals, in addition to fatty oils polyunsaturates of the invention, in order to assist the individual ingestion of the minimum daily requirements of these substances In addition to the polyunsaturated fatty acids listed above, it may also be desirable to supplement the nutritional composition with zinc, copper, and acid. In addition to antioxidants, it is believed that these substances also they will provide an elevation to the stressed immune system and thus provide other benefits to the individual. The presence of zins, on or roughly fuzzy is opsional and is not required in order to gain the beneficial effects of immune suppression. Likewise, a pharmaceutical composition can be supplemented with these same substances. In a more preferred embodiment, the nutritional formula contains, in addition to the antioxidant system and the polyunsaturated fatty acid component, a source of carbohydrate wherein at least 5% by weight of the carbohydrate is an indigestible oligosaccharide. In yet another preferred modality, the nutritional composition additionally contains protein, taurine and carnitine. The polyunsaturated fatty acids, or derivatives thereof, made by the method described can be used as dietary substitutes, or supplements, particularly formulas for lastantee, for patients undergoing intravenous feeding or for preventing or treating malnutrition. Typically, human breast milk has a fatty acid profile ranging from about 0.15% to about 0.36% as docosahexaenoic acid, from about 0.03% to about 0.13% -like eicosapentaenoic acid, from about 0.30% to about 0.88% as arachidonic acid. , from about 0.22% to about 0.67% as dihomo-gamma-linolenic acid, and from about 0.27% to about 1.04% as gamalinolenic acid. Additionally, the predominant triglyceride in human milk has been reported to be 1, 3-di-oleoyl-2-palmitoyl, with 2-palmitoyl glycerides reported as better absorbed than 2-oleoyl or 2-lineoyl glycerides (US Pat. Number: 4,876,107). Thus, fatty acids such as arachidonic acid, dihomo-gamma-linolenic acid, gamalinolenic acid and / or eicosapentaenoic acid produced by the invention can be used to alter the composition of infant formulas to better replicate the poly fatty acid composition - Unsaturated from human breast milk. In particular, a fat composition for use in a pharmacological or food supplement, particularly a breast milk substitute or supplement, will preferably comprise one or more of arachidonic acid, dihomo-gamma-linolenic acid and gamalinolenic acid. More preferably, the oil will comprise from about 0.3 to 30% arachidonic acid, from about 0.2 to 30% dihomo-gamma-linolenide acid, and from about 0.2 to about 30% acetic acid alloy. In addition to the concentration, the proportions of arachidonic acid, dihomo-gamma-linolenic acid and gamalinolenic acid can be adapted for a particular end use. When formulated as a breast milk supplement, or substitute, a fat composition containing two or more of arachidonic acid, dihomo-gamma-linolenic acid and gamalinolenic acid will be provided in a ratio of about 1:19:30 to about 6: 1. : 0.2, respectively. For example, animal breast milk can vary in proportions of arachidonic acid: dihomo-gamma-linolenic acid: gamma-linolenic acid ranging from 1:19:30 to 6: 1: 0.2, which includes intermediate proportions that are preferably about 1: 1: 1, 1: 2: 1, 1: 1: 4. When they are produced together in a host cell, the rate adjustment and the percentage conversion of the precursor substrate such as gamalinolenic acid and dihomo-gamma-linolenic acid into arachidonic acid can be used to accurately control the proportions of polyunsaturated fatty acids. unsaturated For example, a conversion rate of 5% to 10% of dihomo-gamma-linolenic acid in arachidonic acid can be used to produce a ratio of arachidonic acid to dihomo-gamma-linolenic acid of about 1:19, while a Conversion ratio of about 75% to 80% can be used to produce a ratio of arachidonic acid against dihomo-gamma-linolenic acid of about 6: 1.

Therefore, either in a cell culture system or in a host animal, regulation of the time, extent and specificity of desaturase expression as described can be used to modulate the levels and proportions of polyunsaturated fatty acids. . Depending on the expression system used, eg, cell culture or an animal that expresses fats in its milk, the fats can also be isolated and recombined in the concentrations and proportions desired. The amounts of fats provided by these proportions of polyunsaturated fatty acids can be determined following standard protocols. The polyunsaturated fatty acids, or the host cells that contain them, can also be used as animal feed supplements to alter an animal tissue or fatty acid composition of milk to a more desirable one - for human or animal consumption. For dietary supplement, the polyunsaturated fatty acids or their derivatives can be incorporated in cooking fats, oils or margarines formulated so that in normal use the recipient receives the desired sanctity. Polyunsaturated fatty acids can also be incorporated into infant formulas, nutritional supplements or other food products, and may find use as anti-inflammatory or cholesterol lowering substances. Pharmaceutical Compositions The present invention also encompasses a pharmaceutical composition comprising one or more of the resulting acids and / or fats produced according to the methods disclosed therein. More specifically, this pharmaceutic somposision may comprise one or more of the acids and / or fats as well as a standard, well-known, non-toxic pharmaceutically acceptable carrier, adjuvant or carrier such as, for example, phosphate buffered saline, water, ethanol , polyols, vegetable oils, a wetting substance or an emulsion such as water / oil emulsion. The compositions may be in liquid or solid form. For example, the composition may be in the form of a tablet, capsule, liquid or ingestible powder, injectable, or topical ointment or cream. Possible routes of administration include, for example, oral, rectal and parenteral. The route of administration, of course, will depend on the desired effect. For example, if the composition is used to treat rough, dry, or old skin, to treat injured or burned skin, or to treat skin or hair affected by a disease or condition, it may be applied topically. The dose of the composition to be administered to the patient can be determined by someone with ordinary skill in the art and depends on various factors such as the weight of the patient, the age of the patient, the immunological state of the patient, etc. With respect to the form, the composition can be, for example, a solution, a dispersion, a suspension, an emulsion or a sterile powder that is reconstituted. Additionally, the composition of the present invention can be used for cosmetic purposes. It can be added to pre-existing cosmetic compositions so that a mixture is formed or can be used as a composition alone. The pharmaceutical compositions can be used to administer the polyunsaturated fatty acid component to an individual. Suitable pharmaceutical compositions may comprise sterile physiologically acceptable aqueous or non-aqueous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution in sterile solutions or dispersions for ingestion. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, and the like), convenient mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. You can maintain proper fluency, for example, for the maintenance of the required particle size in the case of dispersions and for the use of surfactants. It may be desirable to include isotonic substances, for example sugars, sodium chloride and the like. In addition to these inert diluents, the composition may also include adjuvants, such as wetting substances, emulsifiers and suspending substances, sweeteners, flavoring-and perfuming substances. The suspensions, in addition to the active compounds, may contain suspending substances, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth or mixtures of these substances, and the like. Solid dosage forms such as tablets and capsules can be prepared using techniques well known in the art. For example, the polyunsaturated fatty acids of the invention can be made into tablets with conventional tablet bases such as lactose, sucrose, and corn starch in combination with binders such as asasia, corn starch or gelatin, disintegrating substances such as potato starch or alginic acid and a lubricant such as stearic acid or magnesium stearate. The capsules can be prepared by incorporating these excipients into a gelatin capsule together with antioxidants and the polyunsaturated fatty acid component. The amount of the antioxidants and the polyunsaturated fatty acid component that should be incorporated in the pharmaceutical formulation should be adjusted within the guidelines discussed above. As used in this application the term "treat" refers to either preventing, or reducing the incidence of, the unwanted occurrence. For example, treating immune suppression refers either to avoiding the occurrence of this suppression or to reducing the amount of this suppression. The terms "patient" and "individual" are being used interchangeably and both refer to an animal. The term "animal" as used in this application refers to any warm-blooded mammal including, but not limited to, dogs, humans, monkeys, and apes. As used in the application, the term "approximately" refers to an amount that varies from the range or number established in a reasonable amount depending on the context of use. Any number or numerical range specified in the specification shall be considered to be modified by the term approximately. "Dosage" and "ration" are used interchangeably and refer to the amount of nutritional or pharmaceutical composition ingested by the patient in a single dose and designed to administer effective amounts of antioxidants and structured triglyceride. As will be readily apparent to those skilled in the art, a single dose or ration of the liquid nutritional powder should supply the amount of anti-oxidants and polyunsaturated fatty acids mentioned above. The amount of the dose or ration should be a volume that a typical adult can consume in one sitting. This amount can vary widely depending on the age, weight, sex or medical condition of the patient. However, as a general guideline, a single serving or dose of liquid nutritional product should be considered to encompass a volume of 100 to 600 milliliters, more preferably 125 to 500 milliliters and more preferably 125 to 300 milliliters. The polyunsaturated fatty acids of the present invention can also be added to foods even when supplementation of the diet is not required. For example, the composition can be added to any of its types including, but not limited to margarines, modified butters, cheeses, leshe, yogurt, chocolate, sweets, snacks, salad oils, cosmetics oils, cosmetics, scabs, pessado and drinks. Pharmaceutical applications For pharmacological use (human or veterinary), sompositions are usually administered orally but can be administered by any route through which they can be successfully absorbed., for example, parenterally (i.e. subcutaneously, intramucosally or intravenously), vaginally or vaginally or topically, for example, as an ointment or skin bond. The polyunsaturated fatty acids of the present invention can be administered alone or in combination are a carrier or excipient pharmaceutically aseptable. When available, gelatin sapsules are the preferred form of oral administration. Dietary supplementation, as presented above, can also provide an oral administration route. The unsaturated acids of the present invention can be administered in dissolved forms, or somo salts, esters, amides or prodrugs of the fatty acids. Any farmasuisably asepable salt is overshadowed by the present invention; Especially preferred are the sodium, potassium or lithium salts. Also included are the N-alkyl polyhydroxamine salts, such as N-methyl glusamine, disclosed in the PCT publication WO 96/33155. Preferred esters are ethyl esters. As solid salts, the polyunsaturated fatty acids can also be administered in the form of a tablet. For intravenous administration, the polyunsaturated fatty acids or derivatives thereof can be insorporated in somersial formulations such as somatic intralipids. The fatty acid profile of normal adult plasma ranges from 6.64 to 9.46% of arachidonic acid, 1.45 to 3.11% of dihomo-gamma-linolenide, and 0.02 to 0.08% of gamalinoléniso. These fatty polyunsaturated fats or their metabolism presurers can be administered, either alone or in mixtures are other fatty polyunsaturated fatty acids, to enhance a profile of normal fat in a pasiente normal. When desired, the individual components of the formulas can be individually provided in the form of a kit or kit, for single or multiple use. A typical dose of a partiscular fatty acid is 0.1 milligrams at 20 grams, or up to 100 grams daily, and is preferably 10 milligrams at 1, 2, 5 or 10 grams per day as required, or equivalent molar sanctities of the derived forms thereof. Parenteral nutrient sompositions that range from about 2 to about 30 wt.% Of salted fatty acids such as triglycerides are insulated by the present invention; a somposisance having from about 1 to about 25 per cent by weight of the somatosamine total polyunsaturated fatty acid somatization is preferred (US Pat. No. 5,196,198). Other vitamins, and in particular fat-soluble vitamins such as vitamins A, D, E and L-sarnitine, can optionally be included. When desired, a preservative such as tosorthol can be added, typically to about 0.1% by weight. The resulting pharmacological sompositions may include solvents, dispersions, suspensions or emulsions, sterile or non-aqueous, physiologically acceptable, and sterile powders for their resuspension in sterile solutions or unusable dispersions. Examples of carriers, diluents, solvents or solvents and non-aqueous solvents include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, and the like), mixtures thereof, vegetable products (such as olive oil) and immiscible organoesters. such as ethyl oleate. Proper fluidity can be maintained, for example, by maintaining the required particle size in cases of dispersions and by the use of surfactants. It may also be desirable to include isotonic compositions, for example, sugars, sodium slurium and the like. In addition to these inert diluents, the somposision may also include adjuvants, such as wetting substances, emulsifiers and suspending agents, sweeteners, flavorings and perfumes. The suspensions, in addition to the active compounds, may contain suspensory substances, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances and the like. . An especially preferred pharmaceutical composition contains diacetyltartaric acid esters of mono and diglycerides dissolved in a medium or aqueous solvent. The diacetyltartaric acid esters of mono and diglycerides have a HLB hydrolytic equilibrium value of about 9-12 and are significantly more hydrophilic than existing antimicrobial lipids having HLB values of 2-4. These existing hydrophobic lipids can not be formulated in aqueous compositions. As described in this, the lipids can now be solubilized in aqueous medium in combination with diacetyl tartaric acid esters of mono and diglycerides. According to this embodiment, the diacetyltartaric acid esters of mono and diglycerides (e.g., DATEM-C12: 0) are combined with other asthmatic antimicrobial lipids (e.g., 18: 2 and 12: 0 monoglycerides) and mix to obtain a homogeneous mixture. The homogen allows an increased antimicrobial activity. The mixture can be completely dispersed in water. This is not possible without the addition of diacetyltartaric acid esters of mono and diglycerides and pre-mixing with other monoglycerides before introduction into water. The aqueous composition can be mixed under sterile conditions with physiologically acceptable diluents, preservatives, regulators or propellants as may be required to form a spray or an inhalant. The present invention also encompasses the treatment of numerous disorders with fatty acids. Supplementation with the polyunsaturated fatty acids of the present invention can be used to treat restenosis after angioplasty. Symptoms of inflammation, rheumatoid arthritis, and asthma and psoriasis can be treated with the polyunsaturated fatty acids of the present invention. Evidence indicates that polyunsaturated fatty acids may be involved in calcium metabolism, suggesting that the polyunsaturated fatty acids of the present invention can be used in the treatment or prevention of osteoporosis and kidney or kidney stones. urinary The polyunsaturated fatty acids of the present invention can be used in the treatment of cancer. It has been shown that malignant cells have altered fatty acid compositions; the addition of fatty acids has been shown to decrease their growth and cause cell death, and increase their susceptibility to chemotherapeutic substances. Gamalinolenic acid has been shown to cause re-expression in cancer cells of cellular adhesion molecules E-cadherin, the loss of which is associated with aggressive metastasis. Clinical trials in the intravenous administration of water-soluble lithium salt of gamalinolenic acid to cancer patients in the pancreas produced statistically significant increases in their survival. Supplementation with polyunsaturated fatty acids may also be useful to treat cachexia associated with cancer. The polyunsaturated fatty acids of the present invention can also be used to treat diabetes (U.S. Patent Number: 4,826,877; Horrobin et al., Am. J. Clin. Nutr. Vol. 57 (Suppl.), 732S-737S). Metabolism and composition of altered fatty acids have been demonstrated in diabetic animals. These alterations have suggested that they are involuntary in some of the. long-term sompyslays resulting from diabetes, including retinopathy, neuropathy, nephropathy, and reproductive system damage. Primrose oil, which contains gamalinolenic acid, has been shown to prevent and reverse nerve damage in diabetics. The polyunsaturated fatty acids of the present invention can be used to treat eczema, reduce blood pressure and improve math scores. Deficiency in essential fatty acids has been suggested to be involved in eczema, and some studies have shown beneficial effects on eczema from treatment with gamalinolenic acid. Gamalinolenic acid has also been shown to reduce increases in blood pressure associated with stress, and improve performance in arithmetic tests. Gamalinolenic acid and dihomo-gamma-linolenic acid have been shown to inhibit platelet aggregation, cause vasodilation, lower cholesterol levels, and inhibit smooth muscle proliferation of the vessel wall and fibrous tissue.

(Brenner et al., Adv. Exp. Med. Biol. Vol. 83, p.85-101, 1976). The administration of acetyl-gamalinoliene or dihomo-gamma-linolenase, alone or in combination, is eicosapentaenoic acid, has been shown to reduce or prevent gastrointestinal bleeding and other side effects caused by non-steroidal anti-inflammatory drugs (United States Patent Number: 4,666,701). Gamalinolenic acid and dihomo-gamma-linolenic acid have also been shown to prevent or treat endometriosis and premenstrual syndrome (U.S. Pat. No. 4,758,592) and to treat myalgic encephalomyelitis and chronic fatigue after viral infections. (Patent of the United States of North America Number: 5,116,871). Other uses of the polyunsaturated fatty acids of this invention include the use in treatment of AIDS, multiple sclerosis, acute respiratory syndrome, hypertension and inflammatory skin disorders. The polyunsaturated fatty acids of the invention can also be used for formulas for general health as well as for geriatric treatments. Veterinary applications It should be noted that the farsuchesis and nutrimental disarmament can be used in connection with animals, as well as humans, since animals experience muses of the same needs and sonsions as humans. For example, the sausage or sauces of the present invention can be used in animal feed supplements. The following examples are presented by way of illustration, not limitation. EXAMPLES Example 1. Isolation of a sesuensia of 5-desaturase nusleotides of Mortierella alpina Example 2. Expression of slones 5-desaturase of M. alpina in baker's yeast Example 3. Invisal optimization of sultivo's editions Example 4. Distribusion of Fatty polyunsaturated fatty acids in yeast lipid frassions Example 5. Other optimization of sultives Example 6. Identifisation of homologs of? 5 and? 6 desaturases of M. alpina Example 7. Identification of homologs? 5 and? 6 of M__ alpina in others organisms that produce polyunsaturated fatty acids Example 8. Identification of homologues? 5 and? 6 of M. alpina in other organisms that produce polyunsaturated fatty acids Example 9. Sequences of human desaturase Example 10. Nourishing compositions Example 1 Isolation of a sequence of nucleotides? 5-desaturase from Mortierella alpina Mortierella alpina produces arachidonic acid (ARA, 20: 4) from the precursor 20:30 by a? 5-desaturase. A nucleotide sequence coding for the? 5 desaturase from Mortierella alpina was obtained through polymerase chain reaction amplification using the first strand of the M. alpina cDNA and degenerate oligonucleotide primers corresponding to amino acid sequences conserved between? 6 desaturases of Synechocystis and Spirulina. The procedure used was as follows: The total RNA was isolated from a culture that produces polyunsaturated fatty acids three days old.

Mortierella alpina using the protocol of Hoge et al. (1982), Experimental Mycology 6: 225-232. RNA was used to prepare double-stranded cDNA using the BRL Lambda-ZipLox system, following the manufacturer's instructions. Fractions of various sizes of M. alpina DNAs were impasted separately to produce libraries with inserts of different average size. The "full length" library contains approximately 3 x 10 clones with an average insert size of 1.77 kb. The "sequestration grade" library contains approximately 6 x 10 clones with an average insert size of 1.1 kb. In reverse, 5 micrograms of the total RNA were transcribed using the BRL SuperscriptRTase and the TSyn primer (5'-CCAAGCTTCTGCAGGAGCTCTTTTTTT TTTTTTTT-3 '), SEQ ID NO: 10. Degenerate oligonucleotides were designated to regions conserved between the two? 6 sequences. -cyanobacterial desaturase.

The specific primers used were D6DESAT-F3 (SEQ ID NO: 8) (5'-CUACUACUACUACAYCAYÁCOTAYACOAAYAT-3 ') and D6DESAT-R3 (SEQ ID NO: 9) (5 '-CAUCAUCAUCAU0GGRAA0ARRTGRTG-3'), where Y = C + T, R = A + G, and 0 = I + C. Amplification was carried out by polymerase chain reaction in a 25 microliter volume containing: annealed derivative of 40 nanograms of total RNA, 2 pM of each primer, 200 μM of deoxyribonucleotide triphosphate, 60 mM Tris-Cl, pH 8.5, 15mM (NH4) 2S04, 2 mM MgCl2. The samples were subjected to an initial denaturation step of 95 degrees (all temperatures are Celsius) for 5 minutes, then maintained at 72 degrees while 0.2 U of Taq polymerase was added. The polymerase chain reaction thermocycling conditions were as follows: 94 degrees for 1 minute, 45 degrees-for 1.5 minutes, 72 degrees for 2 minutes. The polymerase chain reaction continued for 35 cycles. The polymerase chain reaction using these primers in the first strand of the M. alpina cDNA produced a reaction product of 550 base pairs. Comparison of the deduced amino acid sequence of the M. alpina polymerase chain reaction fragment SEQ ID NO: 3 revealed regions of homology to? 6-desaturases (see Figure 5). However, there was only about 28% identity over the compared region. The product of the polymerase chain reaction was used as a probe to isolate the corresponding DNA slides from a library of M. alpina. The longest cDNA clone, Ma29, was designated pCGN5521 and sequenced severely in both strains. The cDNA is contained in an insert of 1481 base pairs in the pZL1 vector (Bethesda Research Laboratories) and, beginning as the first ATG, contains an open reading frame that encodes 446 amino acids. The reading frame contains the deduced sequencing of the polymerase chain reaction fragment. The sequence of the cDNA insert was found to contain regions of homology to? 6-desaturases (see Figure 5). For example, it was found that three conserved "histidine boxes" (which had been observed in desaturases attached to the membrane (Okuley et al., (1994) The Plant Cell 6: 147-158)) were found in Mortierella sesuensia in the amino acid positions 171-175, 207-212, and 387-391 (see Figure 3). However, the motif of the typical amino acid "HXXHH" for the histidine saja terrase of the Mortierella desaturase was shown to be QXXHH, SEQ ID NO: 11-12. Surprisingly, the amino terminus of the sodifised protein showed significant homology to sitosromo b5 proteins. In this way, the DNA slit of Mortierella appears to be a fusion between a sitosom b5 and the fatty acid desaturase. Since the cytochrome or b5 is believed to function as an electron donor for the membrane bound desaturases enzymes, it is possible that the N-terminal sitosom b5 domain of this desaturase protein is involved in its function. This may be advantageous when the desaturase is expressed in heterologous systems for the production of polyunsaturated fatty acids. EXAMPLE 2 Expression of M. alpina desaturase clones in baker's yeast Yeast transformation The transformation of lithium asetate of the yeast was carried out in accordance with standard protocols (Methods in Enzymology, Vol. 194, p. 186-187, 1991). In summary, the yeast was grown in yeast peptone (YPD) balance at 30 aC. The cells were centrifuged, resuspended in TE, centrifuged again, resuspended in TE containing 100 mM lithium acetate, centrifuged again, and resuspended in TE / lithium acetate. The resuspended yeast was incubated at 30 ° C for 60 minutes with shaking. The carrier DNA was added and the yeast was divided into tubes. Transformant DNA was added and the tubes were incubated for 30 minutes at 30 ° C. PEG solution (35% (weight on volume) PEG 4000, 100 mM lithium acetate, TE pH7.5) was added followed by 50 minutes of incubation at 30 ° C. A 5 minute salting shoke was carried out at 42 ° C the cells were agglomerated, washed with TE, agglomerated again and resuspended in TE. The resuspended cells were plated on selective media. Expression of desaturases in the transformed yeast The cDNA clones of Mortierella alpina were selected to determine the desaturase activity in the baker's yeast. A? 15-desaturase from canola was used (obtained by polymerase chain reaction using the first cDNA strand from culture seeds 212/86 of Brassica napus using primers based on the published sequence (Arondel et al. Science 258: 1353-1355)) was used as a positive control. The? 15-desaturase gene and the Ma29 cDNA clone gene were inserted into the pYES2 expression vector (Invitrogen), resulting in the plasmids pCGR-2 and pCGR-4, respectively. These plasmids were transfected in yeast strain 334 cerevisiae yeast 334 and were expressed after induction with galactose and in the presence of substrates that allowed the detection of specific desaturase activity. The control strain was strain 334 of S. cerevisiae which contained the unchanged vector pYES2. The substrates used, the products produced and the indicated desaturase activity were: dihomo-gamma-linolenic acid (conversion to arachidonic acid would indicate activity? 5-desaturase), linolenic acid (conversion to gamalinolenic acid would indicate activity? 6-desaturase; in alpha-linolenic acid would indicate activity? 15-desaturase), oleic acid (an endogenous substrate made by S. cerevisiae, conversion to linolenic acid would indicate activity? 12-desaturase, of which S. cerevisiae lacks), or arachidonic acid (the conversion to eicosapentaenoic acid would indicate activity? 17-desaturase). The results are given in Table 1 below. The lipid fractions were extracted as follows: Cultures were grown for 48-52 hours at 15 ° C. The cells were pelleted by centrifugation, washed once with sterile bidistilled water, and re-agglomerated. The agglomerate was centrifuged with methanol; chloroform was added together are tritridecanoin (as an internal standard). The mixtures were incubated for at least 1 hour at room temperature or at 4 ° C overnight. The chloroform layer was extracted and filtered through a Whatman filter with one gram of anhydrous sodium sulfate to remove particles from the wastewater. The organic solvents were evaporated at 40 ° C under a stream of nitrogen. The extracted lipids were derived in methyl esters of fatty acids (EMAG) for gas chromatography (GC) analysis by adding 2 milliliters of 0.5 N potassium hydroxide in methanol to a closed tube. The samples were heated at 95 ° C to 100 ° C for 30 minutes and cooled to room temperature. Approximately 2 milliliters of 14% boron trifluoride in methanol was added and heating was repeated. After the extracted lipid mixture was cooled, 2 milliliters of water and 1 milliliter of hexane were added to extract the methyl esters of fatty acids to be analyzed by gas chromatography. The percentage conversion was calculated by dividing the product produced by the sum of (the product produced and the added substrate) and then multiplied by 100. To sample the percentage conversion of oleic acid, as no substrate was added, the total linolenic acid produced was divided between the sum of (oleic acid and the produced linolenic acid), and then multiplied by 100.

Table 1. Expression of M. alpina Desaturase in Baker's Yeast The slone of sontrol? 15-desaturase exhibited 16.3% substrate sonification. The pCGR-4 slone expressing the Ma29 cDNA gave 15.3% of the substrate 20: 3 at 20: 4? 6, indicating that the gene sodifices a? 5-desaturase. The background (non-specific substrate conversion) was between 0 and 3% in these cases. We also introduce substrate inhibition of the activity using different consentrations of the substrate. When substrate was added at 100 μM, the percent sonotage to the produst was lowered in su parasion to the substrate was added at 25 μM (see below). Additionally, varying the sonsentrasions of the dihomo-gamma-linolenide substrate, from about 5 μM to about 200 μM of porsentual sonification of dihomo-gama-linolenium in arachidonoid acid varied from about 5% to 75% are the? 5-desaturase from M. alpina. These data show that desaturases can be expressed in different substrate species in a heterologous system and used to produce polyunsaturated long-chain fatty acids. Table 2 represents the fatty acids of interest as a percentage of the total lipids extracted from the yeast host S. serevisiae 334 are the indiscriminate plasmid. There was no glusosa present in the medium of sresimiento. Gas affinity chromatography was used to separate the resinous lipids. Gas / MS chromatography was used to verify the identity produtos. The produsto expected for the? 5-desaturase, harsh α-linolense from B. napus, was challenged by sucking its substrate, linolenic acid, exogenously added to the induced yeast culture. This finding demonstrates that the yeast expression of a desaturase gene can produce functional enzyme and detectable amounts of product under ordinary culture conditions. Both exogenously added substrates were collected by yeast, although slightly less than the longer chain polyunsaturated fatty acid, dihomo-linolenic acid (20: 3), was incorporated into the yeast than into the linolenic acid (18: 2) when either was added in free form to the induced yeast cultures. Arachidonic acid was detected as a novel polyunsaturated fatty acid in yeast when dihomo-γ-linolenic acid was added as the substrate to S. cerevisiae 334 (pCGR-4). This identifies pCGR-4 (Ma29) as the? 5-desaturase of M. alpina. Prior to this, the isolation and expression of the 5-desaturase from any source had not been reported.

Table 2 Fatty Acid as Percentage of Total Lipid Extracted from Yeast 100 μM substrate added * 18: 1 is an endogenous fatty acid in yeast Key Table 18: 1 = oleic acid 18: 2 = linolenic acid a-18: 3 = a-linolenic acid? -18: 3 = acid? linolénioo 18: 4 = stearidonic acid 20: 3 = dihomo -? - linolenic acid 20: 4 = arachidonic acid Example 3 Optimization of Culture Conditions Table 3A shows the effect of the exogenous free fatty acid substrate concentration on yeast incorporation and conversion to fatty acid product as a percentage of total extracted yeast lipid. In all cases, low amounts of exogenous substrate (1-10 μM) resulted in a low fatty acid substrate incorporation and product formation. At a concentration between 25 and 50 μM of free fatty acid in the culture and induction medium produced the highest percentage of fatty acid product formed, while the concentration of 100 μM and the high incorporation in yeast yeast subserves decreased or inhibited the desaturase astivity. The inhibition of back-feeding of high fatty acid substrate concentration was well illustrated when the percentage conversion rates of the respective fatty acid substrates to their respective products were compared in Table 3B. In all cases, the substrate consorption 100mM in the sresimiento medium decreased the percentage conversion to the product. The effect of the average composition was also evident when glucose was present in the growth medium for the 5-desaturase, since the substrate absorption in percent decreased by 25 μM (Table 3A). However, the percentage conversion by? 5-desaturase increased by 18% and the porsentual product formed remained the same in the presence of glucose in the growth medium. Table 3A Effect of the Substrate Added on the Percentage of Incorporated Substrate and Product Formed in Yeast Extracts Table 3B Effect of Concentration of the Substrate in the Media on the Percentage of Conversion of Greases in Product in Yeast Extracts to. there is no glusosa in the middle b. Yeast peptone broth (YPA) c. 18: 1 is an endogenous yeast lipid its. is ND substrate concentration not available Table 4 shows the amount of fatty acid produced by a recombinant desaturase from yeast cultures induced when different amounts of free fatty acid substrate are used. The weight of the fatty acid was determined since the total amount of lipid varied dramatically when culture conditions changed, such as the presence of glucose in the yeast growth and the induction medium. To better determine the conditions when the recombinant desaturase would produce more polyunsaturated fatty acid product, the amount of individual fatty acids was examined. The absence of glucose reduced the amount of arachidonic acid produced by? 5-desaturase by almost half. For ? -desaturase the amount of total yeast lipid decreased by almost half in the absence of glucose.

Table 4 Fatty Acid Produced in μM from Yeast Extracts without glucose in the middle of their. is consentrasión of substrate ND (not available) s 18: 1, the substrate, is a lipid of endogenous yeast Example 4 Distribution of polyunsaturated fatty acids in fractions of yeast lipids Table 5 illustrates the insorporasión of free fatty acids in their new prodrugs formed in yeast lipids distributed in the major lipid frassions. A total lipid extract was prepared as described above. The extra lipid was separated in TLC plasmas, and the frassions were identified by shading them are standard. Scratch bands were resoled, and internal standards were added. The frassions were saponified and methylated previously, and subjected to gas chromatography. Gas chromatography salsuló the sanctity of fatty acid somparándolo are a standard. Paraseria that substrates are assesibles in phospholipid form to desaturasas. Table 5 Distribution of Fatty Acid in Various Fractions of Yeast Lipid in μg SC = S. serevisiae (plasmid) Example 5 Additional Culture Optimization The growth and induction conditions for the optimal activities of desaturases in Saccharomyces cerevisiae were evaluated. The different culture conditions that were manipulated for optimal activity were: i) induction temperature, ii) inductor concentration, iii) substrate addition time, iv) substance concentration, v) sugar source, vi) phase of growth in induction. These studies were used using the gene? 5-desaturase from Mortierella alpina (Ma29). In addition, the effect of changing the host strain on the expression of the? 5-desaturase gene was also determined. As described above, the best substrate conversion rate in arachidonic acid was observed at the substrate concentration of 1 μM, however, the percentage of arachidonic acid in the total fatty acids was higher in the substrate sonsension of 25 μM. To determine if the substrate needed to be modified to an easily available form before it could be used by the desaturase, the substrate was added either 15 hours after the induction or concomitant with the addition of the inducer (indicated as below, in Figure 6A ). As can be seen in Figure 6A, the addition of the substrate prior to induction did not have a significant effect on the activity of 5-desaturase. In fact, the addition of the substrate together with the inducer was slightly better for the expression / activity of the 5-desaturase, as the levels of arachidonic acid in the total fatty acids were higher. However, the conversion rate of the substrate into the product was slightly lower. The etiology of the expression inducer concentration / activity of 5-desaturases of Mortierella was examined by inducing SC334 / pCGR5 with 0.5 or 2% (weight / volume) of galactose. As shown in Figures 7A and 7B, the expression of? 5-desaturase was higher when induced with 0.5% galactose. In addition, the conversion rate of the substrate into the product was also better when SC334 / pCGR5 was induced with 0.5% galactose versus 2% galactose. To determine the effect of temperature on the 5-desaturase activity, host strain SC334, transformed with pCGR5 (SC334 / pCGR5) was cultured and induced at 15 ° C, 25 ° C, 30 ° C and 37 ° C. The amount of arachidonic acid (20: 4n6) produced in SC334 / pCGR5 cultures, supplemented with 20: 3n6 substrate, was measured by fatty acid analysis. Figure 8A represents the amount of 20: 3n6 and 20: 4n6, expressed as a percentage in total fatty acids. Figure 8B represents the conversion rate of the substrate into product. The growth and induction of SC334 / pCGR5 at 25 ° C, was the best for the expression of? 5 -desaturase as evidenced by the high levels of arachidonic acid in the total fatty acids.

Additionally, the highest rate of sonification of substrate in produsto was also presented at 25 BC. The sultivo and the induction at 15EC gave the lowest expression of arachididae asid, while at 37 aC it gave the lowest sonotration of substrate in produsto. The efesto of the yeast strain in the expression of the? D-desaturase gene was studied in 5 different host hosts: INVSC1, INVSC2, YTC34, YTC41, and SC334, at 159C and 30aC. At 15aC, the SC334 had the highest percentage of arachidonic acid in total fatty acids, suggesting greater activity of 5-desaturase in SC334. The conversion rate of the substrate into product, however, is lower in SC334 and higher in INVSC1 (Figures 9A and B). At 30 SC, the highest percentage of product (arachidonic acid) in the total fatty acids was observed in INVSC2, although the rate of substrate sonication in produst in INVSC2 was slightly lower than in INVSC1 (Figures 10A and B). The acidic arachidonide, the 5-desaturase prodrug, was kneaded into the phospholipid phosphate (Example 4). Therefore the sanctity of the arid arachidus produced in yeast is limited by the sanctity that can be soured in the phospholipid phosphate. If the harsh arachidonid could also be soured in other frassions such as the triglyceride frassion, the sanctity of the arid arachidus produced in yeast could be increased. • To test this hypothesis, the gene 5-desaturase was expressed in the yeast host strain DBY746 (obtained from the Yeast Genetic Stock Center, 1021 Donner Laboratory, Berkeley, CA 94720. The genotype of strain DBY746 is Mato;, his3-? l, leu2-3, leu2-112, ura3-32, trpl-289, gal). Yeast strain DBY746 has an endogenous gene for choline transferase. The presence of this enzyme could allow the DBY746 strain to convert the excess phospholipids into a triglyceride fraction. The results in Figure 11 show that the conversion of the substrate into product does not increase compared to SC334, which does not have the gene for choline transferase. To study the effect of the medium on the expression of 5-desaturase, pCGR4 / SC334 was cultured in four different media at two different temperatures (15 ° C and 30 ° C) and in two different host strains (SC334 and INVSC1). The composition of the media was as follows: Medium A: mm-Ura, + 2% galactose + 2% glucose. Medium B: mm-Ura, + 20% galactose + 2% glucose + 1 M sorbitol (pH5.8) Medium C: mm-Ura, + 2% galactose + 2% raffinose. Medium D: mm-Ura, + 2% galactose + 2% raffinose + 1 M sorbitol (pH5.8) mm = minimum medium The results show that the highest conversion rate of the substrate in product at 15 ° C in SC334 was observed in medium A. The highest global sonication rate for? 5-desaturase in SC334 was at 30 B in medium D. The highest rate of 5-desaturase sonication in INVSC1 was also at 30a in medium D ( Figures 12A and 12B). These data show that a DNA that sodifises a desaturase that can be converted to a dihomo-gamma-linolenide in an arachidonic acid can be isolated from Mortierella alpina and can be expressed in a heterologous system and used to produce polyunsaturated long chain fatty acids. The production of arachidonic acid is exemplified from the dihomo-gama-linolense acidic presurer by the expression of a? -desaturase in yeast. Example 6: Homologues for the homologues for? 5 and? 6 desaturases of M. alpina A sesuensia of nusleiso was identified that sodifises an? 5-desaturase presumed through the search of TBLASTN from the databases est through NCBI using the amino acids 100-446 of Ma29 somo a sonults. The truncated porsion of the Ma29 sesuensia was used to avoid resoger homologies based on the porosión of sitosromo b5 in the N-terminus of the desaturase. The deduced amino acid sequence of a Dictyostelium dissoideum est (number of asseso C25549) shows a very significant homology are Ma29 and a minor one, but still signifisativa homology, are Ma524. DNA sesuence is presented as SEQ ID NO: 13. The amino acid sesuensia is presented as SEQ ID NO: 14. Example 7 Identification of homologues? 5 and? 6 of M. alpina in other organisms that produce polyunsaturated fatty acids To look for the involuntary desaturases in the production of polyunsaturated fatty acids, a library of DNAs was constructed from total RNA isolated from Phaeodastylum trisornutum. A plasmid-based DNA library was constructed in pSPORTl (GIBCO-BRL) following the instrussions of the manufacturer using a kit or somersially available kit (GIBCO-BRL). Scissors of random DNAs were identified and sesuensias of nusleid acid were identified, which sodifish? 5 or? 6 desaturasas presumed through the BLAST search of the databases and sompasion are the sesuensias of Ma29 and Ma524. A slone from the librarian of the Phaeodastylum was identified and is homologous with Ma29 and Ma524; called 144- 011-B12. The DNA sequence is presented as SEQ ID NO: 15. The amino acid sesuensia is presented as SEQ ID NO: 16. Example 8 Identification of homologues? 5 and? 6 of M. alpina in other organisms that produce polyunsaturated fatty acids To look for the involuntary desaturases in the production of polyunsaturated fatty acids, a DNA library was constructed from RNA isolated from the Sshizoshytrium thickener. A plasmid-based cDNA library was constructed in pSPORTl (GIBCO-BRL) following the manufacturer's instructions using a commerically available set (GIBCO-BRL). Sequences of random DNAs were identified and sesuensias of nusleiso were identified that sodifise? 5 or? 6 desaturasas presumed through the BLAST search of the databases and somparasión with the sequencies of Ma29 and Ma524. A clone was identified from the library Schizoshytrium are homology are Ma29 and Ma524; called 81-23-C7. This clone contained an insert of approximately one kb. The parsial sequencing was obtained from the extreme end of the week using the universal session selectors to the front and back. The DNA sequence from the forward primer is presented as SEQ ID NO: 17. Sequence of peptide is presented as SEQ ID NO: 18. The DNA sesuensia from the reverse primer is presented as SEQ ID NO: 19. The Amino acid sessias of the reverse primer is presented as SEQ ID N0: 20. EXAMPLE 9 Human Desaturase Gene Sequences Human desaturase gene sequences potentially involved in the biosynthesis of long chain polyunsaturated fatty acids were isolated based on the homology between the human DNA sesuensias and the sesuensias of the desaturase gene of Mortierella alpina . The three conserved "histidine sajas" that are known to be preserved between the desaturases attached to the membrane were ensontraron. As are other desaturases attached to the membrane, the final said motif of histidine HXXHH was shown to be QXXHH. The amino acid sesuensia of the presumed human desaturases exhibited homology are the? 5,? 6,? 9, and? L2 desaturases of M. alpina. The sesuensia of Δ5 desaturase and Δ6 desaturase DNA from M. alpina were used to refer to the LifeSeq database of Insyte Pharmaseutisals, Ins. , Palo Alto, California 94304. The ε-desaturase sesuensia was divided into fragments: (1) aminoasids numbers 1-150; (2) amino acid numbers 151-300; and (3) amino acid numbers 301-446. The ε6 desaturase sesuensia was divided into three fragments: (1) aminoasids numbers 1-150; (2) amino acid numbers 151-300; and (3) amino acid numbers 301-457. These polypeptide fragments were investigated using the database using the "tblastn" algorithm. This algorithm compares a sesuensia of sonulta of protein sontra a database of sesuensia of dynamic nusleotides between tradusida in the six lession marsos (both sadenas). The fragments 2 and 3 of the polypeptide of α 5 and α 6 of M. alpina have homologies are the sesuensias of CloneID as presented in Table 6. The CloneID represents a sesuensia - individual from the LifeSeq database of Incyte. After the results of "tblastn" were reviewed, clone information was searched with the defaults reference values of = 50, and Productssore = 100 for different ClonelD numbers. The results of clone information displayed the information that includes ClusterID, CloneID, Library, HitID, Hit Descripsion (hit). When they were selected, the ClusterID number displayed the clone information of all the clones that belonged in that ClusterID. The Asse ble (assemble) command assembles all the CloneIDs that comprise the ClusterID. The following default reference values were used for the GCG assembly (Genetics Computer Group, University of Wisconsin Bioteshnology Center, Madison, Wissonsin 53705): Word size: 7 Minimum overlap: 14 Rigor: 0.8 Minimum identity: 14 Maximum gap: 10 Hole weight: 8 Length weight: 2 GCG assembly results displayed contigs generated based on sequence information within ClonelD. A contig is a DNA sequence alignment based on areas of homology between these sequences. A new sesuensia (sonsense sesuensia) was generated based on the DNA sesuensias aligned in a sontig. The sonnet that was CloneID was identified, and the ambiguous sites of the sonsense sesuensia were edited based on the alignment of the ClonelDs (see SEQ ID NO: 21 - SEQ ID NO: 25) to generate the best possible sesuensia. The rescheduling was repeated for the six ClonelDs listed in Table 6. This produced five unique contigs. The sonsense edits of the 5 sonigs were imported into the software program Sequencher (Gene Codes Corporation, Ann Arbor, Michigan 48 105). These sonsenso sequels were assembled. Sontig 2511785 overlaps is sontig 3506132, and this new sontig was named 2535 (SEQ ID NO: 27). The Sequensher program sonts were inspired by the GCG session analysis software package. Each contig was translated into the six reading frames in protein sequencing. The sesuensia? 5 (Ma29) and? 6 (Ma524) of M. alpina are collected are one of the traduced sontigs using the FastA search (a search by Pearson and Lipman to determine the similarity between a sesultan of sesulta and a group of sequences of the same type (nusleiso acid or protein)). The homology between these sesuencias suggests the open reading frames of sada sontig. The homology between the? 5 and? 6 of M. alpina are the sontigs 2535 and 3854933 were used to srear the final sontig called 253538a.

Figure 13 is the FastA comparison selection of the final contig 253538a and Ma29, and Figure 14 is the FastA comparison selection of the final contig 253538a and Ma524. The DNA sequences for the different contigs are presented in SEQ ID NO: 21-SEQ ID NO: 27. The different peptide sequences are shown in SEQ ID NO: 28-SEQ ID NO: 34. Although the open lesion subarachnoid is generated by combining the two sonigs, contig 2535 shows that there is a single sequence at the start of this contig that does not match with contig 3854933. Therefore, it is possible that these sontigs were generated from independent desaturase or human genes. Contig 253538a contains an open reading frame that encodes 432 amino acids. Start with Gln (CAG) and end with the stop codon (TGA). Contig 253538a is aligned with both sequence? 5 and? 6 of M. alpina, suggesting that it could be either of the two desaturases, as well as other known desaturases that share homology with each other. The individual contigs listed in Table 6, as well as the intermediate contig 2535 and the final sontig 253538a can be used to isolate the complete genes for human desaturases. Uses of human desaturases These human sequences can be expressed in yeasts and plants using the procedures described in the previous examples. For expression in transgenic mammalian and animal cells, these genes can provide a higher codon deviation. These human sequences can also be used to identify related desaturase sequences. Table 6 EXAMPLE 10 Nutrient Compositions The polyunsaturated fatty acids of the above examples can be used in various nutritional supplements, formulations for infants, nutritional substitutes and other solutions for nutrition. I. FORMULATIONS FOR INFANTS A. Isomil® Soy Formula with Iron. Use: As a beverage for infants, children and adults with an allergy or sensitivity to cow's milk. A diet for patients with disorders for which baldness should be avoided: lactase deficiency, lactose intolerance and galactosemia. Features: Soy protein isolate to avoid allergy symptoms or sensitivity to cow's milk protein Formulation without lactose to prevent lactose-associated diarrhea Low osmolality (240 mOsm / kilograms of water) to reduce the risk of osmotic diarrhea. Dual carbohydrates (corn syrup and sucrose) designed to increase the absorption of carbohydrates and reduce the risk of exceeding the absorption capacity of the damaged intestine. 1.8 milligrams of iron (as ferrous sulfate) per 100 salterns to help prevent the defism of iron. Recommended levels of vitamins and minerals. Vegetable oils to provide the recommended levels of esensial fatty acids. Milk white color, milk-like consistency and pleasant aroma. Ingredients: (Pareve, ®) 85% water, 4.9% corn syrup, 2.6% sugar (sucrose), 2.1% soybean oil, 1.9% soy protein isolate, 1.4% coso oil, 0.15% citrate of calcium, 0.11% of tribasic calcium phosphate, potassium citrate, potassium phosphate monobasic, potassium slurry, mono and diglycerides, soy lesitis, sarragenin, assorbic acid, L-methionine, magnesium sulfate, potassium phosphate dibasium, sodium slurium, soline slurium, taurine, ferrous sulfate, m-inositol, alpha-tosoferyl asetate, zinsulfate, L-sarnitine, niasinamide, salty pantothenate, supersaturated sulphate, vitamin A palmitate, thiamine slorhydride , riboflavin, pyridoxine slorohydrate, acid fuchsia, manganese sulfate, potassium iodide, phylloquinone, biotin, sodium selenite, vitamin D3 and sianosobalamin. B. Isomil® DF Soy Formula for Diarrhea. Use: As a long-term diet for dietary management of diarrhea in babies and babies. Faces: First formula for infants to contain added dietary fiber from soy fiber specifically for the management of diarrhea. Clinically it shows to reduce the duration of production of loose, watery stools during mild to severe diarrhea in infants. Nutrimentally complete to meet the nutritional needs of the infant. Soy protein isolate with added L-methionine that meets or exceeds infant requirements of all essential amino acids. Formulation without lactose to prevent diarrhea associated with lactose. Low osmolality (240 mOsm / kilogram of water) to reduce the risk of osmotic diarrhea. Dual carbohydrates (corn syrup and sucrose) designed to increase the absorption of carbohydrate and reduce the risk of exceeding the absorption capacity of the damaged intestine. Satisfies or exceeds the vitamin or mineral levels recommended by the Nutrition Committee of the American Academy of Pediatrics and those required by the Infant Formula Act. 1.8 milligrams of iron (as ferrous sulfate) per 100 calories to help prevent iron deficiency.

Vegetable plants to provide the respected levels of fatty esensial fatty acids. Ingredients: (Pareve, ®) 86% water, 4.8% corn syrup, 2.5% azusar (sasarose), 2.1% soybean meal, 2.0% soy protein isolate, 1.4% bland diet, 0.77% fiber of soy, 0.12% of sodium salt, 0.11% of phosphate of tribasic acid, 0.10% of potassium sitrate, potassium chloride, potassium phosphate monobásiso, mono and diglycerides, soy lesitis, sarragenin, magnesium sulfate, assiduous acid , L-methionine, potassium phosphate dibasium, sodium slurium, soline slurry, taurine, ferrous sulfate, m-inositol, alpha-tocopheryl asetate, zinc sulfate, L-sarnitine, niasinamide, pantothenium salt, supersaturated sulfate, vitamin A palmitate, thiamin chloride slorhydrate, riboflavin, • pyridoxine hydrochloride, acid fuchsin, manganese sulfate, potassium iodide, phylloquinone, biotin, sodium selenite, vitamin D3 and sianosobalamin. C. Isomil® SF Soy Formula without Sucrose with Iron Usage: As a drink for mothers, children and adults are an allergy or sensitivity to the protein of vasa leshe or an intolerance to sucrose. A feeding for pasantes are disorders for the suals should be avoided the ballast and the sucrose. Carasteristisas: Soy protein isolate to avoid allergy symptoms or sensitivity to cow's milk protein. Formulation without balm to prevent diarrhea associated with lactose. Sucrose free for the patient who can not tolerate sucrose. Low osmolality (180 Osm / kilogram of water) to reduce the risk of osmotic diarrhea. 1.8 milligrams of iron (as ferrous sulfate) per 100 calories to help prevent iron deficiency. Recommended levels of vitamins and minerals. Vegetable oils to provide the recommended levels of essential fatty acids. White milk color, milk-like consistency and pleasant aroma. Ingredients: (Pareve, ®) 75% water, 11.8% hydrolyzed corn starch, 4.1% isolated from soy protein isolate, 2.8% coconut oil, 1.0% modified corn starch, 0.38% tribasic calcium phosphate , 0.17% potassium citrate, 0.13% potassium chloride, mono and diglycerides, soy lecithin, magnesium chloride, ascorbic acid, L-methionine, calcium carbonate, sodium chloride, choline chloride, carrageen, taurine, sulfate ferrous, m-inositol, alpha-tocopheryl acetate, zinc sulfate, L-carnitine, niacinamide, calcium pantothenate, cupric sulfate, vitamin A palmitate, thiamine chloride hydrochloride, riboflavin, pyridoxine slorohydrate, rough fuchsia, sulfate of manganese, potassium iodide, phylloquinone, biotin, sodium selenite, vitamin D3 and cyanocobalamin. D. Isomil® 20 Soybean with Iron formula ready for food, 20 calories / per 29.6 milliliters Use: When you want a diet with soy. Ingredients: (Pareve, ®) 85% water, 4.9% corn syrup, 2.6% sugar (sucrose), 2.1% soybean oil, 1.9% soy protein isolate, 1.4% cauliflower oil, 0.15% salsium sitrate, 0.11 % of tribasic salsium phosphate, potassium sitrate, potassium phosphate monobaside, potassium slurry, mono and diglycerides, soy lesitis, sarragenin, asbestos acid, L-methionine, magnesium sulfate, dibasic potassium phosphate, sodium chloride, choline chloride, taurine, ferrous sulfate, m-inositol, alpha-tosoferyl acetate, zins sulfate, L-sarnitine, niasinamide, pantothenium salt, supersaturated sulphate, vitamin A palmitate, thiamine slorhydride, riboflavin, slorohydrate of pyridoxine, acid fuchsia, manganese sulfate, potassium iodide, phylloquinone, biotin, sodium selenite, vitamin D3 and sianosobalamin. E. Similac® Formula for infants Use: When a formula is needed for lactation: if the decision is taken to de-sodium feed before the age of one year, if a supplementation is needed - breastfeeding or feeding routine if breast feeding is not adopted. Characteristics: Protein of quality and adequate quantity for the good development; denatured by heat, which reduces the risk of enteric blood loss associated with milk. Grease from a mixture of vegetable oils (doubly homogenized), which provides essential linoleic acid that is easily absorbed. Carbohydrate as lactose in a proportion similar to that of human milk Low renal solute load to minimize stress on the organs that develop. Forms in powder, liquid consented and ready to be taken. Ingredients: (®-D) water, skim milk, lactose, soybean oil, coconut oil, mono and diglycerides, soy lecithin, ascorbic acid, carrageenan, choline chloride, taurine, m-inositol, alpha-tocopheryl acetate , zinc sulfate, niacinamide, ferrous sulfate, calcium pantothenate, cupric sulfate, vitamin A palmitate, thiamine chloride hydrochloride, riboflavin, pyridoxine hydrochloride, folic acid, manganese sulfate, phylloquinone, biotin, sodium selenite, vitamin D3 and cyanocobalamin. F. Yes ilac® NeoCare Formula with iron for premature infants Use: For the special nutritional needs of the premature infant after leaving the hospital. Similac NeoCare is a nutrimentally complete formula developed to provide premature infants with the extra calories, proteins, vitamins and minerals needed to promote weight gain and support their development. Characteristics: Reduces the need for caloric and vitamin supplementation. More calories (22 calories / per 29.6 milliliters) than formulas for the standard term (20 calories per 29.6 milliliters). Mixture of highly absorbed fats, with medium chain triglycerides (MCT oil) to help meet the special digestive needs of premature infants. High levels of proteins, vitamins and minerals per 100 calories to extend the nutritional support started in the hospital. More calcium and phosphorus to improve the mineralization of the bones. Ingredients: ®-D corn syrup solids, defatted milk, lactose, milk whey protein concentrate, soybean oil, high oleic acid oleagnet oil content, fractionated coconut oil (medium chain triglycerides), oil coconut, potassium citrate, tribasic calcium phosphate, calcium carbonate, ascorbic acid, magnesium chloride, potassium chloride, sodium chloride, taurine, ferrous sulfate, m-inositol, choline chloride, ascorbyl palmitate, L-carnitine , alpha-tocopheryl acetate, zinc sulfate, niacinamide, mixed tocopherols, sodium citrate, calcium pantothenate, cupric sulfate, thiamine slurium hydrochloride, vitamin A palmitate, beta sarotene, riboflavin, pyridoxine slorohydrate, folic acid, Manganese sulfate, phylloquinone, biotin, sodium selenite, vitamin D3 and cyanocobalamin. G. Si ilac Natural Fortifier of human milk low in iron of natural care ready for use, 24 calories / 29.6 milliliters. Use: Designed to be mixed with human milk or to be fed alternately with human milk for low birth weight infants. Ingredients: ®-D water, skim milk, hydrolyzed corn starch, lactose, fractionated coconut oil (medium chain triglycerides), whey protein concentrate, soybean oil, coconut oil, tribasic calcium phosphate, citrate of potassium, magnesium chloride, sodium citrate, ascorbic acid, calcium carbonate, mono and diglycerides, soy lecithin, carrageen, choline chloride, m-inositol, taurine, niacinamide, L-carnitine, alpha-tocopheryl acetate, sulphate zinc, potassium chloride, calcium pantothenate, ferrous sulfate, cupric sulfate, riboflavin, vitamin A palmitate, thiamine chloride hydrochloride, pyridoxine slorohydrate, biotin, rough fuchsia, manganese sulfate, phylloquinone, vitamin D3, selenite sodium and cyanocobalamin. Various polyunsaturated fatty acids of this invention can be substituted and / or added to the infant formulas described above and to other infant formulas known to those skilled in the art. II. NUTRITIVE FORMULATIONS A. ENSURE® Use: ENSURE is a low-residue waste liquid food primarily designed as an oral nutrient supplement to be used with or between meals or, in adequate quantities to replace a meal. ENSURE is free of lactose and gluten, and is suitable for use in modified diets, including low-cholesterol diets. Although it is mainly an oral supplement, it can be fed by tube. Patient Conditions: For patients on modified diets For geriatric patients are nutrition risk For patients with involuntary weight loss For patients recovering from illness or surgery For patients who need a low-residue diet Ingredients: ®-D Water, sugar ( sucrose), maltodextrin (corn), calcium and sodium caseinates, high oleic oil, safflower oil, soy protein isolate, soybean oil, canola oil, potassium citrate, tribasic calcium phosphate, sodium citrate , magnesium chloride, dibasic magnesium phosphate, artificial flavor, sodium chloride, soy lecithin, choline chloride, ascorbic acid, carrageenan, zinc sulfate, ferrous sulfate, alpha-tocopheryl acetate, gelan gum, niacinamide, pantothenate of calcium, manganese sulfate, cupric sulfate, vitamin A palmitate, thiamine chloride hydrochloride, pyridoxine hydrochloride, riboflavin, phobic acid lico, sodium molybdate, chromium chloride, biotin, potassium iodide, sodium selenate. B. ENSURE® BARS Use: ENSURE BARRAS is a balanced nutrition, complete for supplementary use between or with the somidas. It provides a delicious, nutrient-rich alternative to other snacks. ENSURE BARS contains less than 1 gram of lactose / per bar, and Brownie flavor of chosolate and gluten-free. (The Honey Graham Crunsh flavor is gluten). Patient Conditions: For people who need extra salories, proteins, vitamins and minerals. Especially useful for people who do not take enough calories and nutrients. For people who have the ability to chew and swallow. It can not be used by people with allergies to peanuts or any type of allergy. the nuts . Ingredients: Honey Graham Crunch - Corn syrup high in fruit content, isolated from soy protein, brown sugar, honey, maltodextrin (corn), puffed rice (ground rice, sugar [sucrose], salt [sodium chloride]. and malt), oat bran, partially hydrogenated soybean and cottonseed oils, soy polysaccharide, glycerin, whey protein concentrate, polydextrose, fructose, calcium caseinate, cosoa powder, artificial flavors, oil cañola, high safflower oil in oleic content, defatted dry milk, whey powder, soy lecithin and corn oil. Manufactured in facilities that process nuts. Vitamins and minerals: Tribasic calcium phosphate, dibasic potassium phosphate, magnesium oxide, salt (sodium chloride), potassium chloride, ascorbic acid, ferric orthophosphate, alpha-tocopheryl acetate, niacinamide, zinc oxide, calcium pantothenate , copper gluconate, manganese sulfate, riboflavin, beta-carotene, pyridoxine hydrochloride, thiamine mononitrate, folic acid, biotin, chromium chloride, potassium iodide, sodium selenate, sodium molybdate, phylloquinone, vitamin D3 and cyanocobalamin . Protein: Honey Graham Crunch - The protein source is a mixture of soy protein isolate and milk proteins. Soy protein isolate 74% milk proteins 26% Fat: Honey Graham Crunch - The source of fat is a mixture of partially hydrogenated oils of cottonseed and soybeans, canola oils (rape seed), high safflower content of oleic, and corn and soy lecithin. Seed oils of partially hydrogenated corn and soybeans. 76% Canola oil 8% Safflower oil high in oil 8% Corn stew 4% Soy lecithin 4% Carbohydrates: Honey Graham Crunch - The source of carbohydrates is a combination of high fructose corn syrup, brown sugar, maltodextrin , honey, puffed rice, glycerin, soy polysaccharide, and oat bran. High fructose corn syrup 24% Brown sugar 21% Maltodextrin 12% Honey 11% Inflated rice 9% Glycerin 9% Soy polysaccharide 7% Bran. oatmeal 7% C. ENSURE® HIGH PROTEIN Use: ENSURE HIGH PROTEIN is a high protein liquid food concentrate designed for people who require additional calories, proteins and vitamins and minerals in their diets. It can be used as a nutritional supplement orally or between meals or in adequate amounts as a meal replacement. ENSURE HIGH PROTEIN is free of lactose and gluten, and is suitable for use by people recovering from general surgery or hip fractures and by patients at risk of pressure ulcers. Patient Condition For patients who require additional calories, proteins, vitamins and minerals, such as patients recovering from general surgery or hip fractures, patients are at risk of pressure ulcers, and patients on low-cholesterol diets. Characteristics Low in saturated fat Contains 6 grams of total fat and less than 5 milligrams of cholesterol per trace. Taste creamy risot Excellent source of protein, calcium, and other essential vitamins and minerals For low cholesterol diets Free of lactose, easily digested Ingredients: Supreme Vanilla: Water ®-D, sugar (sasarose), maltodextrin (corn), sodium and sodium saseinates, high oleic acid safflower, soy protein isolate, soybean oil, canola oil, sodium Potassium, Triassic Salsium Phosphate, Sodium Sitrate, Magnesium Chloride, Dibasic Magnesium Phosphate, artificial flavor, sodium slurry, soy lesion, solin slurry, ascorbic acid, carrageenan, zinc sulfate, ferrous sulfate, alpha-tosoferyl acetate, Gellan gum, niasinamide, salty pantothenate, manganese sulfate, sulfate on, vitamin A palmitate, thiamine sloride slorhydrate, pyridoxine slorhydrate, riboflavin, acid fuchsin, sodium molybdate, chromium slurium, biotin, potassium iodide, sodium selenate, phylloquinone, vitamin D3 and cyanocobalamin. Protein The protein source is a mixture of two proteins of high biological value: casein and soy. Sodium and Calsium Caseinates 85% Soy Protein Isolate 15% Fat: The source of fat is a mixture of three sauces: high oleic oleifera, cañola, and soybean high oleic safflower oil 40% Canola oil 30 % Soy oil 30% The level of fat in ENSURE HIGH PROTEIN satisfies the American Heart Association (AHA) guidelines. The 6 grams of fat in ENSURE HIGH PROTEIN represent 24% of total calories, with 2.6% of fat coming from saturated fatty acids and 7.9% of polyunsaturated fatty acids. These values are within the lineaments of the Amerisan Heart Assosiation de <30% of total fat salories, < 10% salidos of saturated fatty acids, and < 10% of total salories of polyunsaturated fatty acids. Carbohydrate: ENSURE HIGH PROTEIN are a combination of maltodextrin and sasarose. The soft sweetness and the variety of flavors (supreme of vanilla, shosolate royal, wild blackberry -and banana), plus VARI-FLAVORSO® Flavor Pacs in pecan, cherry, strawberry, lemon and orange help to avoid taste fatigue and help Satisfy the patient's taste. Vanilla and other flavors without chocolate Sucrose 60% Maltodextrin 40% Chocolate Sucrose 70% Maltodextrin 30% D. ENSURE® LIGHT Use: ENSURE LIGHT is a low-fat liquid food designed to be used as an oral nutritional supplement with or between rolls. ENSURE LIGHT is free of gluten and gluten, and is suitable for use in modified diets, including low cholesterol diets. Patient Conditions: For normal-weight or past-weight patients who need extra nutrition in a supplement that contains 50% less fat and 20% fewer calories than ENSURE. For healthy adults who do not eat properly and need extra nutrition. Characteristics: Low in fats and saturated fats Contains 3 grams of total fat per serving and < 5 milligrams of cholesterol Taste riso, samoso Exselente source of salsio and other vitamins and minerals esensiales For diets low in cholesterol Without lactose, easily digested Ingredients: French vanilla: Water ® -D, maltodextrin (corn), azusar (sasarose), salsio saseinate, high sapham aseptic in oyster medium, sanala, magnesium chloride, sodium citrate, potassium citrate, potassium phosphate dibasium, magnesium phosphate dibasium, natural flavor and Artifisial, Triassium Salsium Phosphate, Cellulose Gel, Soline Chloride, Soya Lesitin, Sarragenin, Salt (Sodium Slurry), Ascorbic Acid, Cellulose Gum, Ferrous Sulfate, Alpha-Tocopheryl Sulfate, Zinsulfate, niasinamide, manganese sulfate, salsium pantothenate, supersaturated sulfate, thiamine sloride slorhydrate, vitamin A palmitate, pyridoxine slorhydrate, riboflavin, sódico slurium, acid fuchsia, sodium molybdate, biotin, potassium iodide, sodium selenate , phylloquinone, vitamin D3 and sianosobalamin. Proteins: The source of protein is salsium saseinate Caseinate 100% Salty Fat The source of fat is a mixture of two sauces: high safflower in oleic content and canola. High oleic safflower oil 70% Canola oil 30% The level of fat in ENSURE LIGHT satisfies the lineaments of the American Heart Association (AHA). The 3 grams of fat in ENSURE LIGHT represent 13.5% of the total calories, being 1.4% of the fat of saturated fatty acids and 2.6% of polyunsaturated fatty acids. These values are within the guidelines of the American Heart Association of = 30% of total calories from fat, < 10% of calories from saturated fatty acids, and = 10% of total calories of polyunsaturated fatty acids. ENSURE LIGHT Carbohydrate contains a combination of 'maltodextrin and sucrose. The chosolate flavor is also corn syrup. The soft sweetness and the variety of flavors (vanilla to the fransesa, supreme chocolate, strawberry milkshake), plus VARI-FLAVORS® Flavor Pacs of pecan, cherry, strawberry, lemon and orange, help to avoid flavor fatigue and favor patient cooperation. Vanilla and other flavors that are not chocolate Sucrose 51% Maltodextrin 49% Chocolate Sucrose 47.0% Corn syrup 26.5% Maltodextrin 26.5% Vitamins and Minerals A serving of 236.80 milliliters of ENSURE LIGHT provides at least 25% of the RDI of 24 vitamins and minerals keys Caffeine The chocolate flavor contains 2.1 milligrams of caffeine per 236.80 milliliters. E. ENSURE PLUS® Use: ENSURE PLUS is a low-residue, high-calorie liquid food for use when extra nutrients and calories are needed but a normal protein concentration. It is primarily designed as an oral nutritional supplement to be used with or between foods or, in adequate amounts, as a meal replacement. ENSURE PLUS does not have lactose or gluten. Although it is mainly an oral nutritional supplement, it can be fed by tube. Patient Conditions: For patients requiring extra calories and nutrients, but a normal concentration of protein, in a limited volume For patients who need to gain or maintain healthy weight Characteristics Rich, creamy flavor Good source of essential vitamins and minerals Ingredients Vanilla: Water ® -D, corn syrup, maltodextrin (corn), corn oil, sodium and calcium caseinates, sugar (sucrose), soy protein isolate, magnesium chloride, potassium citrate, tribasic calcium phosphate, soy lecithin, natural and artificial flavor, sodium citrate , potassium chloride, choline chloride, ascorbic acid, carrageenan, zinc sulfate, ferrous sulfate, alpha-tocopheryl acetate, niacinamide, calcium pantothenate, manganese sulfate, cupric sulfate, thiamine chloride hydrochloride, pyridoxine hydrochloride, riboflavin, vitamin A palmitate, folic acid, biotin, chromium chloride, sodium molybdate, potassium iodide, sodium selenite, phylloquinone, cyanocobalamin and vitamin D3. Protein The protein source is a mixture of two proteins of high biological value: casein and soy. Sodium and calcium caseinates 84% Soy protein isolate 16% Fat The source of fat is corn oil. Corn oil '100% Carbohydrate ENSURE PLUS contains a combination of maltodextrin and sucrose. The soft sweetness and the variety of flavors (vanilla, chocolate, strawberry, coffee, pecan cream and egg shake), plus VARI-FLAVORS® Flavor Pacs of pecan, cherry, strawberry, lemon and orange, help to avoid taste fatigue and help patient cooperation . Flavors of vanilla, strawberry, pecan cream, and coffee Corn honey 39% Maltodextrin 38% Sucrose 23% Flavors chocolate and egg shake Corn syrup 36% Maltodextrin 34% Sucrose 30% Vitamins and minerals A serving of 236.80 milliliters of fluid ENSURE PLUS provides at least 15% of the RDI of 25 key vitamins and minerals Caffeine per chocolate contains 3.1 milligrams of caffeine per 236.80 milliliters. The brown flavor contains a clean amount of caffeine. F. ENSURE PLUS® HN Use: ENSURE PLUS HN is a nutritionally complete high calorie, high nitrogen content liquid food designed for people with high calorie and protein needs or limited volume tolerance. It can be used for oral supplement or for total nutritional support per tube. ENSURE PLUS HN lacks lactose and gluten. Patient Conditions: For patients with increasing calorie and protein needs, such as after surgery or injury For patients with limited volume tolerance and early satiety Characteristics For supplemental or total nutrition For oral or tube feeding 1.5 CaVmL High nitrogen Calorically dense Ingredients: Vanilla: Water ®-D, maltodextrin (corn), sodium and calcium caseinates, corn stew, azusar (sasarose), soy protein isolate, magnesium chloride, potassium citrate, tribasic calcium phosphate, soy lecithin, natural and artificial flavor, sodium citrate, choline chloride, ascorbic acid, taurine, L-carnitine, zinc sulfate, ferrous sulfate, alpha-tocopheryl acetate, niacinamide, carrageen, calsium pantothenate, manganese sulfate, cupric sulfate, thiamine chloride hydrochloride, pyridoxine hydrochloride, riboflavin, vitamin A palmitate, acid fuchsin, biotin, chromium chloride, molibdat or of sodium, potassium iodide, sodium selenite, phylloquinone, cyanosobalamin and vitamin D3. G ENSURE® POWDER Use: ENSURE POWDER (reconstituted with water) is a liquid food with low residue content designed primarily as an oral nutritional supplement to be used with or between meals. ENSURE POWDER lacks lactose and gluten, and is suitable for use in modified diets, including low cholesterol diets. Patient conditions: For patients on modified diets For geriatric patients at risk of nutrition For patients recovering from disease / surgery For patients who need a low-residue diet Characteristics Convenient, easy to mix Low in saturated fat Contains 9 grams of total fat and < 5 milligrams of cholesterol per serving High vitamin and mineral content For low cholesterol diets Without lactose, easily digested Ingredients: Vanilla: ®-D corn syrup, maltodextrin (corn), sugar (sucrose), corn oil, caseinates sodium and calcium, isolated from soy protein, artificial flavor, potassium citrate, magnesium chloride, sodium citrate, tribasic calcium phosphate, potassium chloride, soy lecithin, ascorbic acid, choline chloride, zinc sulfate, ferrous sulfate, alpha-tocopheryl acetate, niacinamide, salsium pantothenate, manganese sulfate, thiamine chloride hydrochloride, cupric sulfate, pyridoxine hydrochloride, riboflavin, vitamin A palmitate, fuchsin acid, biotin, sodium molybdate, sodium chloride, chromium, potassium iodide, sodium selenite, phylloquinone, and vitamin D3 and cyanosobalamin. Protein The protein source is a mixture of two proteins of high biological value: saseine and soy. Sodium and calcium caseinates 84% Soy protein isolate 16% Fat The source of fat is corn oil. 100% Corn Oil Carbohydrate ENSURE POWDER contains a combination of corn syrup, maltodextrin, and sucrose. The soft sweetness of ENSURE POWDER, plus VARI-FLAVORS® Flavor Pacs in pasana, sereza, strawberry, lemon, and orange, helps to avoid taste fatigue and favors sooperasión of the pasiente. Vanilla Corn syrup 35% Maltodextrin 35% Sasarosa 30% H. ENSURE® BUDÍN OR NATILLA Use: ENSURE BUDÍN O NATILLA is a dense nutrient supplement that provides balanced nutrition in non-liquid form to be used are or between somites. It is adesuado for diets are sonsistensia odifisada (for example, soft, mashed, or completely liquid) or for people are problems swallowing. ENSURE BUDÍN O NATILLA has no gluten. Conditions of the Patient: For patients are diets of modified sonsistensia (e.g., soft, pureed, or completely liquid) For passions are problems to swallow Characteristics Taste riso and creamy, good Good source of essential vitamins and minerals. Convenient: no refrigeration needed Gluten-free Nutrient profile for 148.00 milliliters: Calories 250, protein 10.9%, total fat 34.9%, carbohydrates 54.2% Ingredients: Vanilla: Leshe skimmed ® -D, water, azúsar (sucrose), parsially hydrogenated soya beans, modified food starch, magnesium sulfate, stearoyl sodium starylate, sodium dibasic phosphate, artifisial taste, assorbic acid, sulphate of zins, ferrous sulfate, alpha-tosoferyl asetate, slurry of choline, niacinamide, manganese sulfate, calsium pantothenate, yellow number 5 FD &C, potassium sitrate, supersaturated sulphate, vitamin A palmitate, thiamine sloride slorhydrate, pyridoxine slorohydrate, riboflavin, yellow number 6 FD &C, rough fuchsin, biotin, phylloquinone, vitamin D3 and sianosobalamin. Protein The protein source is leshe dessremada .. Leshe dessremada 100% Fat The source of fat is aseite of hydrogenated soybeans Hydrogenated soybean 100% Carbohydrate ENSURE NATILLA are a combination of sasarose and modified starch alimentary. The mild sweetness and the variety of flavor (vanilla, shosolate, butter, and tapioca) helps to avoid taste fatigue. The produsto contains 9.2 grams of lactose per serving. Vanilla and other flavors that are not chocolate Sucrose 56% Lactose 27% Modified food starch 17% Chocolate Sucrose 58% Lactose 26% Modified food starch 16% I. ENSURE® WITH FIBER Use: ENSURE WITH FIBER is a nutrimentally somplet liquid food that They are fiber designed for people who can benefit from increased dietary fiber and nutrients. ENSURE WITH FIBER is suitable for people who do not require a low-residue diet. It can be administered orally or by tube, and a nutrient supplement can be used to a regular diet or, in adequate amounts, as a meal replacement. ENSURE WITH FIBER has no lactose or gluten, and is suitable for use in modified diets, including low-cholesterol diets. Patient Conditions For patients who can benefit from increased fiber and dietary nutrients Characteristics New advanced formula low in saturated fat, and higher in vitamins and minerals Contains 6 grams of total fat and < 5 milligrams of cholesterol per serving Taste rich, samoso Good source of fiber Excellent source of vitamins and essential minerals For low-cholesterol diets Without lactose or gluten Ingredients Vanilla: ®-D Water, maltodextrin (corn), sugar (sucrose), calcium and sodium cahenates, oat fiber, high oleic oil safflower oil, canola oil , isolated from soy protein, corn oil, soy fiber, tribasic calcium phosphate, magnesium chloride, potassium citrate, cellulose gel, soy lecithin, dibasic potassium phosphate, sodium citrate, natural and artificial flavors, choline chloride, magnesium phosphate, ascorbic acid, cellulose gum, potassium chloride, carrageenan, ferrous sulfate, alpha-tocopheryl acetate, zinc sulfate, niacinamide, manganese sulfate, calcium pantothenate, cupric sulfate, vitamin palmitate A, thiamine chloride hydrochloride, pyridoxine hydrochloride, riboflavin, folic acid, chromium chloride, biotin, sodium molybdate, potassium iodide, sodium selenate, phylloquinone, vitamin D3 and cyanosobalamin. Protein The protein source is a mixture of two proteins of high biological value, casein and soy. Sodium and Calcium Caseinates 80% Soy Protein Isolate 20% Fat The fat source is a mixture of three oils: high oleic content, cañola and corn. High oleic safflower oil 40% 40% healthy oil 40% cornseed The level of fat in ENSURE WITH FIBER satisfies the lineaments of the Amerisan Heart Association (AHA). The 6 grams of fat in ENSURE WITH FIBER represent 22% of the total calories, with 2.01% of the fat of the saturated fatty acids and 6.7% of the polyunsaturated fatty acids. These values are within the lineaments of the American Heart Association of = 30% of total fat sallorums, <10% of calories from saturated fatty acids, and = 10% of total calories from polyunsaturated fatty acids. Carbohydrate ENSURE WITH FIBER contains a cotnbinasión of maltodextrin and sucrose. The soft sweetness and the variety of flavor (vanilla, chocolate, and pecan cream), plus VARI FLAVORS® Flavor Pacs in pecan, cherry, strawberry, lemon and orange, help to avoid flavor fatigue and favor the cooperation of the owner . Vanilla flavor and others that are not chocolate Maltodextrin 66% Sasarosa 25% Oat fiber 7% Soy fiber 2% Chocolate Maltodextrin 55% Sasarosa 36% Oat fiber 7% Soy fiber 2% Fiber The fiber mixture used in ENSURE WITH FIBER they were in oat fibers and soy polysaccharide. This mixture results in approximately 4 grams of total dietary fiber per can of 236.80 milliliters. The propulsion of insoluble to soluble fiber is 95: 5. The different nutrient supplements previously described and used for others are experiences in teas and can be substituted and / or supplemented by the polyunsaturated fatty acids of this invention. J. Oxepa® Nutrient Product Oxepa is a nutrient-rich, nutrient-rich nutritional product, low in sarbohydrates, designed for dietary management of pastes. they are ARDS or are a risk of ARDS.

-It has an exclusive combination of ingredients, which include a mixture of patented oils containing eicosapentaenoic acid (AEP from fish oil), gamma-linolenic acid (AGL of borage oil), and high levels of anti-oxidants. Caloric distribution: The caloric density is high at 1.5 Cal / mL (355 Cal / 236.80 milliliters), to minimize the volume required to meet energy needs. The distribution of calories in Oxepa is shown in Table 7.

Fat: Oxepa contains 22.2 grams of fat per serving of 236.80 milliliters (93.7 grams per liter). The source of fat is a mixture of oils 31.8% cane oil, 25% medium chain triglycerides (MCTs), 20% borage oil, 20% fish oil, and 3. 2% soy lecithin. The typical fatty acid profile of Oxepa is shown in Table 8. Oxepa provides a balanced amount of monounsaturated and saturated fatty acids, as shown in Table 10. Medium chain triglycerides (MCTs) -.25% of the fat blend - aids in emptying gastric because they are absorbed in the intestinal tract without emulsification by bile acids. The various fatty acid components of Oxepa® nutrient product can be substituted and / or supplemented by the polyunsaturated fatty acids of this invention.

* Fatty acids equal approximately 95% of total fats.

Carbohydrates: The carbohydrate content is 25.0 grams per serving of 236.80 milliliters (105.5 grams per liter). The sources of carbohydrate are 45% maltodextrin (a complex carbohydrate) and 55% sucrose (a simple sugar), both easily digested and absorbed. Oxepa's high fat and low carbohydrate content is designed to minimize the production of carbon dioxide (C02). High levels of C02 can - complicate the disconnection in ventilator-dependent patients. The low level of carbohydrates may also be useful for patients who have developed hyperglycemia induced by tension. Oxepa does not have lactose. Dietary carbohydrate, amino acids from protein, and the glycerol fraction of fats can be converted into glucose within the body. Through this process, the carbohydrate requirements of glucose-dependent tissues (such as the central nervous system and red blood cells) are met. However, a diet without carbohydrates can lead to ketosis, excessive satabolism of tissue protein, and loss of fluid and electrolytes. These efestos can be avoided by the daily ingestion of 50 to 100 grams of digestible carbohydrate, if the caloric intake is adequate. The level of carbohydrates in Oxepa is also sufficient to minimize gluconeogenesis, if energy needs are being met. Oxepa protein contains 14.8 grams of protein per serving of 236. 80 milliliters (62.5 grams per liter). The ratio of calorie / total nitrogen (150: 1) satisfies the need of stressed patients. Oxepa provides enough protein to promote anabolism and maintain lean body mass without precipitating respiratory problems. High protein intakes are a concern in patients with respiratory insufficiency. Although the protein has an effect on the production of carbon dioxide, a diet high in protein will increase the ventilatory problem. The Oxepa protein sources are 86.8% sodium caseinate and 13.2% salsium saseinate. All the publications and patent applications mentioned in this specification are indicative of the level of experience of the technicians in the field to which this invention refers. All publications and patent applications are hereby incorporated by reference to the same extent as if each individual publication or patent application specifically and individually was indicated as incorporated by reference. Having fully described the invention, it will be apparent to a technician with ordinary experience in the field that many changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

LIST OF SEQUENCES (1) GENERAL INFORMATION (i) APPLICANT: Calgene LLC and Abbott Laboratories (ii) TITLE OF THE INVENTION: METHODS AND COMPOSITIONS FOR THE SYNTHESIS OF POLY-INSATURED FATTY ACIDS OF LONG CHAIN • (iii) NUMBER OF SEQUENCES: 34 (2) INFORMATION FOR SEQ ID NO: l: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1483 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: l: GCTTCCTCCA GTTCATCCTC CATTTCGCCA CCTGCATTCT TTACGACCGT TAAGCAAGAT 60 GGGAACGGAC CAAGGAAAAA CCTTCACCTG GGAAGAGCTG GCGGCCCATA ACACCAAGGA 120 CGACCTACTC TTGGCCATCC GCGGCAGGGT GTACGATGTC ACAAAGTTCT TGAGCCGCCA 180 TCCTGGTGGA GTGGACACTC TCCTGCTCGG AGCTGGCCGA GATGTTACTC CGGTCTTTGA 240 GATGTATCAC GCGTTTGGGG CTGCAGATGC CATTATGAAG AAGTACTATG TCGGTACACT 300 GGTCTCGAAT GAGCTGCCCA TCTTCCCGGA GCCAACGGTG TTCCACAAAA CCATCAAGAC 360 GAGAGTCGAG GGCTACTTTA CGGATCGGAA CATTGATCCC AAGAATAGAC CAGAGATCTG 420 GGGACGATAC GCTCTTATCT TTGGATCCTT GATCGCTTCC TACTACGCGC AGCTCTTTGT 480 GCCTTTCGTT GTCGAACGCA CATGGCTTCA GGTGGTGTTT GCAATCATCA TGGGATTTGC 540 GTGCGCACAA GTCGGACTCA ACCCTCTTCA TGATGCGTCT CACTTTTCAG TGACCCACAA 600 CCCCACTGTC TGGAAGATTC TGGGAGCCAC GCACGACTTT TTCAACGGAG CATCGTACCT 660 GGTGTGGATG TACCAACATA TGCTCGGCCA TCACCCCTAC ACCAACATTG CTGGAGCAGA 720 TCCCGACGTG TCGACGTCTG AGCCCGATGT TCGTCGTATC AAGCCCAACC AAAAGTGGTT 780 TGTCAACCAC ATCAACCAGC ACATGTTTGT TCCTTTCCTG TACGGACTGC TGGCGTTCAA 840 GGTGCGCATT CAGGACATCA ACATTTTGTA CTTTGTCAAG ACCAATGACG CTATTCGTGT 900 CAATCCCATC TCGACATGGC ACACTGTGAT GTTCTGGGGC GGCAAGGCTT TCTTTGTCTG 960 GTATCGCCTG ATTGTTCCCC TGCAGTATCT GCCCCTGGGC AAGGTGCTGC TCTTGTTCAC 1020 GGTCGCGGAC ATGGTGTCGT CTTACTGGCT GGCGCTGACC TTCCAGGCGA ACCACGTTGT 1080 TGAGGAAGTT CAGTGGCCGT TGCCTGACGA GAACGGGATC ATCCAAAAGG ACTGGGCAGC 1140 TATGCAGGTC GAGACTACGC AGGATTACGC ACACGATTCG CACCTCTGGA CCAGCATCAC 1200 TGGCAGCTTG AACTACCAGG CTGTGCACCA TCTGTTCCCC AACGTGTCGC AGCACCATTA 1260 TCCCGATATT CTG GCCATCA TCAAGAACAC CTGCAGCGAG TACAAGGTTC CATACCTTGT 1320 CAAGGATACG TTTTGGCAAG CATTTGCTTC ACATTTGGAG CACTTGCGTG TTCTTGGACT 1380 CCGTCCCAAG GAAGAGTAGA AGAAAAAAAAG CGCCGAATGA AGTATTGCCC CCTTTTTCTC 1440 CAAGAATGGC AAAAGGAGAT CAAGTGGACA TTCTCTATGA AGA 1483 (2) INFORMATION FOR SEQ ID NO: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 446 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 2: Met Gly Thr Asp Gln Gly Lys Thr Phe Thr Trp Glu Glu Leu Wing Ala 1 5 10 15 His Asn Thr Lys Asp Asp Leu Leu Leu Wing He Arg Gly Arg Val Tyr 20 25 30 Asp Val Thr Lys Phe Leu Ser Arg His Pro Gly Gly Val Asp Thr Leu 35 40 45 Leu Leu Gly Wing Gly Arg Asp Val Thr Pro Val Phe Glu Met Tyr His 50 55 60 Ala Phe Gly Ala Ala Asp Ala lie Met Lys Lys Tyr Tyr Val Gly Thr 65 70 75 80 Leu Val Ser Asn Glu Leu Pro He Phe Pro Glu Pro Thr Val Phe His 85 85 95 Lys Thr He Lys Thr Arg Val Glu Gly Tyr Phe Thr Asp Arg Asn He 100 105 lio Asp Pro Lys Asn Arg Pro Glu He Trp Gly Arg Tyr Ala Leu He Phe 115, 120 125 Gly Ser Leu He Ala Ser Tyr Tyr Ala Gln Leu Phe Val Pro Phe Val 130 135 140 Val Glu Arg Thr Trp Leu Gln Val Val Phe Ala He He Met Met Gly Phe 145 150 155 160 Ala Cys Ala Gln Val Gly Leu Asn Pro Leu His Asp Ala Ser His Phe 165 170 175 Ser Val Thr His Aßn Pro Thr Val Trp Lys He Leu Gly Wing Thr His 180 185 190 Asp Phe Phe Asn Gly Ala Ser Tyr Leu Val Trp Met Tyr Gln His Met 195 200 205 Leu Gly His His Pro Tyr Thr Asn He Wing Gly Ala Asp Pro Asp Val 210 215 220 Ser Thr Ser Glu Pro Asp Val Arg Arg He Lys Pro Asn Gln Lys Trp 225 230 235 240 Phe Val Asn His He Asn Gln His Met Phe Val Pro Phe Leu Tyr Gly 245 250 255 Leu Leu Wing Phe Lys Val Arg He Gln Asp He Asn He Leu Tyr Phe 260 265 270 Val Lys Thr Asn Asp Wing He Arg Val Asn Pro He Ser Thr Trp His 275 280 '285 Thr Val Met Phe Trp Gly Gly Lys Wing Phe Phe Val Trp Tyr Arg Leu 290 295 300 He Val Pro Leu Gln Tyr Leu Pro Leu Gly Lys Val Leu Leu Leu Phe 305 310 315 320 Thr Val Wing Asp Met Val Being Ser Tyr Trp Leu Wing Leu Thr Phe Gln 325 330 335 Wing Asn His Val Val Glu Val Gln Glp Trp Pro Leu Pro Aslu Glu Asn 340 345 350 Gly He He Gln Lys Asp Trp Wing Ala Met Gln Val Glu Thr Thr Gln 355 360 365 Asp Tyr Ala His Asp Ser His Leu Trp Thr Ser He Thr Gly Ser Leu 370 375 380 Asn Tyr Gln Ala Val His His Leu Phe Pro Asn Val Ser Gln His His 385 390 395 400 Tyr Pro Asp He Leu Wing He He Lys Asn Thr Cys Ser Glu Tyr Lys 405 410 415 Val Pro Tyr Leu Val Lys Asp 'Thr Phe Trp Gln Wing Phe Wing His 420 425 430 Leu Glu His Leu Arg Val Leu Gly Leu Arg Pro Lys Glu Glu 435 440 445 (2) INFORMATION FOR SEQ ID NO: 3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 186 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) ) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 3 Leu His His Thr Tyr Thr Asn He Wing Gly Wing Asp Pro Asp Val Ser 1 5 10 15 Thr Ser Glu Pro Asp Val Arg Arg He Lys Pro Asn Gln Lys Trp Phe 20 25 30 Val Asn His He Asn Gln His Met Phe Val Pro Phe Leu Tyr Gly Leu 35 40 45 Leu Ala Phe Lys Val Arg He Gln Asp He Asn He Leu Tyr Phe Val 50 55 60 Lys Thr Asn Asp Wing He Arg Val Asn Pro He Ser Thr Trp His Thr 65 70 75 80 Val Met Phe Trp Gly Gly Lys Wing Phe Phe Val Trp Tyr Arg Leu He 85 90 95 Val Pro Leu Gln Tyr Leu Pro Leu Gly Lys Val Leu Leu Leu Phe Thr 100 105 110 Val Wing Asp Met Val Ser Ser Tyr Trp Leu Wing Leu Thr Phe Gln Wing 115 120 125 Asn Tyr Val Val Glu Val Val Gln Trp Pro Leu Pro Asp Glu Asn Gly 130 '135 140 He He Gln Lys Asp Trp Ala Ala Met Gln Val Glu Thr Thr Gln Asp 145 150 155 160 Tyr Ala His Asp Ser His Leu Trp Thr Ser He Thr Gly Ser Leu Asn 165 170 175 Tyr Gln Xaa Val His His Leu Phe Pro His 180 185 (2) INFORMATION FOR SEQ ID NO: 4: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 457 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 4 Met Ala Ala Ala Pro Ser Val Arg Thr Phe Thr Arg Ala Glu Val Leu 1 5 10 15 Asn Ala Glu Ala Leu Asn Glu Gly Lys Lys Asp Wing Glu Wing Pro Phe 20 25 '30 Leu Met He He Asp Asn Lys Val Tyr Asp Val Arg Glu Phe Val Pro 35 40 45 Asp His Pro Gly Gly Ser Val He Leu Thr His Val Gly Lys Asp Gly 50 55 60 Thr Asp Val Phe Asp Thr Phe His Pro Glu Ala Wing Trp Glu Thr Leu 65 70 75 80 Wing Asn 'Phe Tyr Val Gly Asp He Asp Glu As Asp Arg Asp He Lys 85 90 95 Asn Asp Asp Phe Wing Wing Glu Val Arg Lys Leu Arg Thr Leu Phe Gln 100 105 110 Ser Leu Gly Tyr Tyr Asp Ser Ser Lys Wing Tyr Tyr Wing Phe Lys Val 115 120 125 Ser Phe Asn Leu Cys He Trp Gly Leu Ser Thr Val He Val Ala Lys 130 135 140 rp Gly Gln Thr Ser Thr Leu Ala Asn Val Leu Ser Ala Ala Leu Leu 145 150 155 160 Gly Leu Phe Trp Gln Gln Cys Gly Trp Leu Wing His Asp Phe Leu His 165 170 175 His Gln Val Phe Gln Asp Arg Phe Trp Gly Asp Leu Phe Gly Wing Phe 180, 185 190 Leu Gly Val Cys Gln Gly Phe Ser Ser Ser Trp Trp Lys Asp Lys 195 200 205 His Asn Thr His His Wing Ala Pro Asn Val His Val Glu Asp Pro Asp 210 215 220 He Asp Thr His Pro Leu Leu Thr Trp Ser Glu His Ala Leu Glu Met 225 230 235 240 Phe Ser Asp Val Pro Asp Glu Glu Leu Thr Arg Met Trp Ser Arg Phe 245 250 255 Met Val Leu Asn Gln Thr Trp Phe Tyr Phe Pro He Leu Ser Phe Wing 260 265 270 Arg Leu Ser Trp Cys Leu Gln Ser He Leu Phe Val Leu Pro Asn Gly 275 280 285 Gln Ala Hiß Lys Pro Ser Gly Ala Arg Val Pro He Ser Leu Val Glu 290 295 300 Gln Leu Ser Leu Ala Met His Trp Thr Trp Tyr Leu Ala Thr Met Phe 305 310 315 320 Leu Phe He Lys Asp Pro Val Asn Met Leu Val Tyr Phe Leu Val Ser 325 330 335 Gln Wing Val Cys Gly Asn Leu Leu Wing He Val Phe Ser Leu Asn His 340 345 350 Asn Gly Met Pro Val He Ser Lyß Glu Glu Wing Val Asp Met Asp Phe 355 360 365 Phe Thr Lys Gln He He Thr Gly Arg Asp Val His Pro Gly Leu Phe 370 375 380 Wing Asn Trp Phe Thr Gly Gly Leu Asn Tyr Gln He Glu His His Leu 385 390 395 400 Phe Pro Ser Met Pro Arg His Asn Phe Ser Lys He Gln Pro Wing Val 405 410 415 Glu Thr Leu Cys Lys Lys Tyr Asn Val Arg Tyr His Thr Thr Gly Met 420 425 430 He Glu Gly Thr Ala Glu Val Phe Ser Arg Leu Asn Glu Val Ser Lys 435 440 445 Ala Ala Ser Lys Met Gly Lys Ala Gln 450 455 ( 2) INFORMATION FOR SEQ ID NO: 5: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 446 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 5 Met Ala Ala Gln He Lys Lys Tyr He Thr Ser Asp Glu Leu Lys Asn 1 5 10 15 His Asp Lys Pro Gly Asp Leu Trp He Ser He Gln Gly Lys Wing Tyr 20 25 30 Asp Val Ser Asp Trp Val Lys Asp His Pro Gly Gly Ser Phe Pro Leu 35 40 45 Lys Ser Leu Wing Gly Gln Glu Val Thr Asp Wing Phe Val Ala Phe His 50 55 60 Pro Wing Ser Thr Trp Lys Asn Leu Asp Lys Phe Phe Thr Gly Tyr Tyr 65 70 75 80 Leu Lys Asp Tyr Ser Val Ser Glu Val Ser Lys Val Tyr Arg Lys Leu 85 90 95 Val Phe Glu Phe Ser Lys Met Gly Leu Tyr Asp Lys Lys Gly His He 100 105 310 Met Phe Ala Thr Leu Cys Phe He Ala Met Leu Phe Ala Met Ser Val 115 120 125 Tyr Gly Val Leu Phe Cys Glu Gly Val Leu Val His Leu Phe Ser Gly 130 135 140 Cys Leu Met Gly Phe Leu Trp He Gln Ser Gly Trp He Gly His Asp 145 150 155 160 Wing Gly His Tyr Met Val Val Ser Asp Ser Arg Leu Asn Lys Phe Met 165 170 175 Gly He Phe Wing Wing Asn Cys Leu Ser Gly He Ser He Gly Trp Trp 180 185 190 Lys Trp Asn His Asn Wing His His Wing Wing Cys Asn Ser Leu Glu Tyr 195 200 205 Asp Pro Asp Leu Gln Tyr He Pro Phe Leu Val Val Ser Ser Lys Phe 210 215 220 Phe Gly Se.r Leu Thr Ser His Phe Tyr Glu Lys Arg Leu Thr Phe Asp 225 230 235 240 Being Leu Being Arg Phe Phe Val Being Tyr Gln His Trp Thr Phe Tyr Pro 245 250 255 He Met Cys Ala Ala Arg Leu Asn Met Tyr Val Gln Ser Leu He Met 260 265 270 Leu Leu Thr Lys Arg Asn Val Ser Tyr Arg Ala Gln Glu Leu Leu Gly 275 280 285 Cys Leu Val Phe Ser He Trp Tyr Pro Leu Leu Val Ser Cys Leu Pro 290 295 300 Asn Trp Gly Glu Arg He Met Phe Val He Wing Ser Leu Ser Val Thr 305 310 315 320 Gly Met Gln Gln Val Gln Phe Ser Leu Asn His Phe Ser Ser Ser Val 325 330 335 Tyr Val Gly Lys Pro Lys Gly Asn Asn Trp Phe Glu Lys Gln Thr Asp 340 345 350 Gly Thr Leu Asp He Ser Cys Pro Pro Trp Met Asp Trp Phe His Gly 355 360 365 Gly Leu Gln Phe Gln He Glu His His Leu Phe Pro Lys Met Pro Arg 370 375 380 Cys Asn Leu Arg Lys He Ser Pro Tyr Val He Glu Leu Cys Lys Lys 385 390 395 400 His Asn Leu Pro Tyr Asn Tyr Ala Ser Phe Ser Lys Ala Asn Glu Met 405 410 415 Thr Leu Arg Thr Leu Arg Asn Thr Wing Leu Gln Wing Arg Asp He Thr 420 425 430 Lys Pro Leu Pro Lys Asn Leu Val Trp Glu Wing Leu His Thr 435 440 445 (2) INFORMATION FOR SEQ ID NO: 6: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 359 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: l ineal ( ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 6 Met Leu Thr Ala Glu Arg He Lys Phe Thr Gln Lys Arg Gly Phe Arg 1 5 10 15 Arg Val Leu Asn Gln Arg Val Asp Wing Tyr Phe Wing Glu His Gly Leu 20 25 30 Thr Gln Arg Asp Asn Pro Ser Met Tyr Leu Lys Thr Leu He He Val 35 40 45 Leu Trp Leu Phe Ser Wing Trp Wing Phe Val Leu Phe Ala Pro Val He 50 55 60 Phe Pro Val Arg Leu Leu Gly Cys Met Val Leu Ala He Ala Leu Ala 65 70 75 80 Wing Phe Ser Phe Asn Val Gly His Asp Wing Asn His Asn Wing Tyr Ser 85 90 95 Ser Asn Pro His As Asn Arg Val Leu Gly Met Thr Tyr Asp Phe Val 100 105 110 Gly Leu Ser Ser Phe Leu Trp Arg Tyr Arg His Asn Tyr Leu His His 115 120 125 Thr Tyr Thr Asn He Leu Gly His Asp Val Glu He His Gly Asp Gly 130 135 140 Wing Val Arg Met Ser Pro Glu Gln Glu His Val Gly He Tyr Arg Phe 145 150 155 160 Gln Gln Phe Tyr He Trp Gly Leu Tyr Leu Phe He Pro Phe Tyr Trp 165 170 175 Phe Leu Tyr Asp Val Tyr Leu Val Leu Asn Lys Gly Lys Tyr His Asp 180 185 190 His Lys He Pro Pro Phe Gln Pro Leu Glu Leu Wing Ser Leu Leu Gly 195 200 205 He Lys Leu Leu Trp Leu Gly Tyr Val Phe Gly Leu Pro Leu Ala Leu 210 215 220 Gly Phe Ser He Pro Glu Val Leu He Gly Wing Ser Val Thr Tyr Met 225 230 235 240 Thr Tyr Gly He Val Val Cys Thr He Phe Met Leu Ala His Val Leu 245 250 255 Glu Ser Thr Glu Phe Leu Thr Pro Asp Gly Glu Ser Gly Wing He Asp 260 265 270 Asp Glu Trp Wing He Cys Gln He Arg Thr Thr Wing Asn Phe Wing Thr 275 280 285 Asn Asn Pro Phe Trp Asn Trp Phe Cys Gly Gly Leu Asn His Gln Val 290 295 300 Thr His His Leu Phe Pro Asn He Cys His He His Tyr Pro Gln Leu 305 310 315 320 Glu Asn He He Lys Asp Val Cys Gln Glu Phe Gly Val Glu Tyr Lys 325 330 335 Val Tyr Pro Thr Phe Lys Wing Wing He Wing Ser Asn Tyr Arg Trp Leu 340 345 350 Glu Wing Met Gly Lys Wing Ser 355 (2) INFORMATION FOR SEQ ID NO: 7: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 365 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 7 Met Thr Ser Thr Thr Ser Lys Val Thr Phe Gly Lys Ser He Gly Phe 1 5 10 15 Arg Lys Glu Leu Asn Arg Arg Val Asn Wing Tyr Leu Glu Wing Glu Asn 20 25 30 He Ser Pro Arg Asp Asn Pro Pro Met Tyr Leu Lys Thr Wing He He 35 40 45 Leu Wing Trp Val Val Ser Wing Trp Thr Phe Val Val Phe Gly Pro Asp 50 55 60 Val Leu Trp Met Lys Leu Leu Gly Cys He Val Leu Gly Phe Gly Val 65 70 75 80 Be Wing Val Gly Phe Asn He Ser His Asp Gly Asn His Gly Gly Tyr 85 90 95 Ser Lys Tyr Gln Trp Val Asn Tyr Leu Ser Gly Leu Thr His Asp Wing 100 105 not He Gly Val Ser Ser Tyr Leu Trp Lys Phe Arg His Asn Val Leu His 115 120 125 His Thr Tyr Thr Asn He Leu Gly His Asp Val Glu He His Gly Asp 130 135 140 Glu Leu Val Arg Met Ser Pro Pro Met Glu Tyr Arg Trp Tyr His Arg 1 5 150 155 160 Tyr Gln His Trp Phe He Trp Phe Val Tyr Pro Phe He Pro Tyr Tyr 165 170 175 Trp Ser He Wing Asp Val Gln Thr Met Leu Phe Lys Arg Gln Tyr His 180 185 190 Asp His Glu He Pro Ser Pro Thr Trp Val Asp He Wing Thr Leu Leu 195 200 205 Wing Phe Lys Wing Phe Gly Val Wing Val Phe Leu He He Pro He Wing 210 215 220 Val Gly Tyr Ser Pro Leu Glu Wing Val He Gly Wing Ser He Val Tyr 225 230 235 240 Met Thr His Gly Leu Val Wing Cys Val Val Phe Met Leu Wing His Val 245 250 255 He Glu. Pro Wing Glu Phe Leu Asp Pro Asp Asn Leu His He Asp Asp 260 »265 270 Glu Trp Wing Wing Gln Val Lys Thr Thr Val Asp Phe Wing Pro Asn 275 280 285 Asn Thr He He Asn Trp Tyr Val Gly Gly Leu Asn Tyr Gln Thr Val 290 295 300 His His Leu Phe Pro His He Cys His He His Tyr Pro Lys He Ala 305 310 315 320 Pro He Leu Wing Glu Val Cys Glu Glu Phe Gly Val Asn Tyr Ala Val 325 330 335 His Gln Thr Phe Phe Gly Ala Leu Ala Wing Asn Tyr Ser Trp Leu Lys 340 345 350 Lys Met Ser He Asn Pro Glu Thr Lys Ala He Glu Gln 355 360 365 [2) INFORMATION FOR SEQ ID NO: 8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 32 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (A) DESCRIPTION: / desc = "Synthetic oligonucleotide (ix) CHARACTERISTICS: (A) NAME / KEY: misc_feature (B) LOCATION: 21 (D) OTHER INFORMATION: / number = 1 / note = "N = Inosine or Cytosine" (ix) CHARACTERISTICS: (A) NAME / KEY: misc_feature (B) LOCATION: 27 (D) OTHER INFORMATION: / number = 2 / note = "N = Inosine or Cytosine" ( xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO 8: CUACUACUAC UACAYCAYAC NTAYACNAAY AT 3 (2) INFORMATION FOR SEQ ID NO: 9: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 27 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (A) DESCRIPTION: / desc = "Synthetic oligonucleotide (ix) CHARACTERISTICS: (A) NAME / KEY: misc_feature (B) LOCATION: 13 (D) OTHER INFORMATION: / number = 1 / note- "N = Inosine or Cytosine" (ix) CHARACTERISTICS: (A) NAME / KEY: misc_feature (B) LOCATION: 19 (D) OTHER INFORMATION: / number = 2 / note = "N = Inosine or Cytosine" ( xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 9: CAUCAUCAUC AUNGGRAANA RRTGRTG (2) INFORMATION FOR SEQ ID NO: 10: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 35 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 10 CCAAGCTTCT GCAGGAGCTC TTTTTTTTTT TTTTT (2) INFORMATION FOR SEQ ID NO: 11: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: aminoaside. (C) CHAIN TYPE: not relevant (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 11 His Xaa Xaa His His l 5 (2) INFORMATION FOR SEQ ID NO: 12: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 12: Gln Xaa Xaa His His 1 5 (2) INFORMATION FOR SEQ ID NO: 13: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 746 nucleic acids (B) TYPE: nucleic acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 13: CGTATGTCAC TCCATTCCAA ACTCGTTCAT GGTATCATAA ATATCAACAC ATTTACGCTC 60 CACTCCTCTA TGGTATTTAC ACACTCAAAT ATCGTACTCA AGATTGGGAA GCTTTTGTAA 120 AGGATGGTAA AAATGGTGCA ATTCGTGTTA GTGTCGCCAC AAATTTCGAT AAGGCCGCTT 180 ACGTCATTGG TAAATTGTCT TTTGTTTTCT TCCGTTTCAT CCTTCCACTC CGTTATCATA 240 GCTTTACAGA TTTAATTTGT TATTTCCTCA TTGCTGAATT CGTCTTTGGT TGGTATCTCA 300 CAATTAATTT CCAAGTTAGT CATGTCGCTG AAGATCTCAA ATTCTTTGCT ACCCCTGAAA 360 GACCAGATGA ACCATCTCAA ATCAATGAAG ATTGGGCAAT CCTTCAACTT AAAACTACTC 420 AAGATTATGG TCATGGTTCA CTCCTTTGTA CCTTTTTTTAG TGGTTCTTTA AATCATCAAG 480 TTGTTCATCA TTTATTCCCA TCAATTGCTC AAGATTTCTA CCCACAACTT GTACCAATTG 540 TAAAAGAAGT TTGTAAAGAA CATAACATTA CTTACCACAT TAAACCAAAC TTCACTGAAG 600 CTATTATGTC ACACATTAAT TACCTTTACA AAATGGGTAA TGATCCAGAT TATGTTAAAA 660 AACCATTAGC CTCAAAAGAT GATTAAATGA AATAACTTAA AAACCAATTA TTTACTTTTG 720 ACAAACAGTA ATATTAATAA ATACAA 746 (2) INFORMATION FOR SEQ ID NO: 14: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 227 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 14 Tyr Val Thr Pro Phe Gln Thr Arg Ser Trp Tyr His Lys Tyr Gln 1 5 10 15 His He Tyr Wing Pro Leu Leu Tyr Gly He Tyr Thr Leu Lys Tyr 20 25 30 Arg Thr Gln Asp Trp Glu Wing Phe Val Lys Asp Gly Lys Asn hey 35 40 45 Wing He Arg Val Ser Val Wing Thr Asn Phe Asp Lys Wing Wing Tyr 50 55 60 Val He Gly Lys Leu Ser Phe Val Phe Phe Arg Phe He Leu Pro 65 70 75 Leu Arg Tyr His Ser Phe Thr Asp Leu He Cys Tyr Phe Leu He 80 85 90 Wing Glu Phe Val Phe Gly Trp Tyr Leu Thr He Asn Phe Gln Val 95 100 105 Ser His Val Wing Glu Asp Leu Lys Phe Phe Wing Thr Pro Glu Arg 110 115 120 Pro Asp Glu Pro Ser Gln He Asn Glu Asp Trp Wing He Leu Gln 125 130 135 Leu Lys Thr Thr Gln Asp Tyr Gly His Gly Ser Leu Leu Cys Thr 140 145 150 Phe Phe Ser Gly Ser Leu Asn His Gln Val Val His His Leu Phe 155 160 165 Pro Ser He Ala Lys Glu Val Cys Asn Phe Thr Glu Met Gly Asn Asp Asp Asp Xaa (2) INFORMATION FOR SEQ ID NO: 15: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 494 nucleic acids (B) TYPE: nucleic acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 15: TTTTGGAAGG NTCCAAGTTN ACCACGGANT NGGCAAGTTN ACGGGGCGGA AANCGGTTTT 60 CCCCCCAAGC CTTTTGTCGA CTGOTTCTGT GGTGGCTTCC AGTACCAAGT CGACCACCAC 120 TTATTCCCCA GCCTGCCCCG ACACAATCTG GCCAAGACAC ACGCACTGGT CGAATCGTTC 180 TGCAAGGAGT GGGGTsTCCA GTACCACGAA GCCGACCTCC TOGACGGGAC CATGCAAOTC 240 TTOCACCATT TGGGCAGCGT GGCCGGCGAA TTCGTCGTGG ATTTTGTACG CGACGGACCC 300 GCCATGTAAT CGTCGTTCGT GACGATGCAA GGGTTCACGC ACATCTACAC ACACTCACTC 360 ACACAACTAG TGTAACTCGT ATAGAATTCG GTGTCGACCT GGACCTTOTT TGACTGGTTG 420 GGGATAGGGT AGGTAGGCGG ACGCGTGGGT CGNCCCCGGG AATTCTGTGA CCGGTACCTG 480 GCCCGCGTNA AAGT 94 (2) INFORMATION FOR SEQ ID NO: 16: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 87 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 16 Phe Trp Lys Xaa Pro Ser Xaa Pro Arg Xaa Xaa Gln Val Xaa Gly 1 5 10 15 Wing Glu Xaa Gly Phe Pro Pro Lys Pro Phe Val Asp Trp Phe Cys 20 25 30 Gly Gly Phe Gln Tyr Gln Val Asp His His Leu Phe Pro Ser Leu 35 40 45 Pro Arg His Asn Leu Wing Lys Thr His Wing Leu Val Glu Ser Phe 50 55 60 Cys Lys Glu Trp Gly Val Gln Tyr His Glu Ala Aßp Leu Val Asp 65 70 75 Gly Thr Met Glu Val Leu Hiß His Leu Gly Ser Val Wing Gly Glu 65 70 75 Phe Val Val Aßp Phe Val Arg Asp Gly Pro Wing Met 80 85 (2) INFORMATION FOR SEQ ID NO: 17: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 520 nucleic acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear ( ii) TYPE OF MOLECULE: nucleic acid (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID N0: 17: GGATGGAGTT CGTCTGGATC GCTGTGCGCT ACGCGACGTG sTTTAAGCGT CATGGGTsCG 60 CTTGGGTACA CGCCGGGGCA GTCGTTGGGC ATGTACTTGT GCGCCTTTGG TCTCGGCTGC 120 ATTTACATTT TTCTGCAGTT CGCCGTAAGT CACACCCATT TGCCCGTGAG CAACCCGGAG 180 GATCAGCTGC ATTGGCTCGA GTACGCGCGG ACCACACTGT GAACATCAGC ACCAAGTCGT 240 GGTTTGTCAC ATGGTGGATG TCGAACCTCA ACTTTCAGAT CGAGCACCAC CTTTTCCCCA 300 CGGCGCCCCA GTTCCGTTTC AAGGAGATCA GCCCGCGCGT CGAGGCCCTC TTCAAGCGCC 360 ACGGTCTCCC TTACTACGAC ATGCCCTACA cGAscsccGT CTCCACCACC TTTGCCAACC 420 TCTACTCCGT CGGCCATTCC GTCGGCGACG CCAAGCsCGA CTAGCCTCTT TTCCTAGACC 480 TTAATTCCCC ACCCCACCCC ATGTTCTOTC TTCCTCCCGC 520 (2) INFORMATION FOR SEQ ID NO: 18: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 153 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: peptide (XÍ) DESCRIPTION OF SEQUENCE: SEQ ID NO: 18 Met Glu Phe Val Trp He Wing Val Arg Tyr Wing Thr Trp Phe Lys 1 5 10 15 Arg His Gly Cys Wing Trp Val His Wing Gly Wing Val Val Gly His 20 25 30 Val Leu Val Arg Leu Trp Ser Arg Leu His Leu His Phe Ser Ala 35 40 45 Val Arg Arg Lys Ser His Pro Phe Wing Arg Olu Oln Pro Gly Gly 50 55 60 Be Ala Ala Leu Ala Arg Val Arg Ala Asp His Thr Val Asn He 65 70 75 Be Thr Lys Ser Trp Phe Val Thr Trp Trp Met Ser Asn Leu Asn 80 85 90 Phe Gln He Olu His His Leu Phe Pro Thr Ala Pro Gln Phe Arg 95 100 105 Phe Lys Glu He Ser Pro Arg Val Glu Ala Leu Phe Lys Arg His 110 115 120 Gly Leu Pro Tyr Tyr Asp Met Pro Tyr Thr Ser Wing Val Ser Thr 125 130 135 Thr Phe Wing Asn Leu Tyr Ser Val Gly His Ser Val sly Asp Wing 140 145 150 Lys Arg Asp \ 2) INFORMATION FOR SEQ ID NO: 19: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 429 nucleic acids (B) TYPE: nucleic acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: nucleic acid (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 19 ACGCGTCCGC CCACGCGTCC GCCGCGAGCA ACTCATCAAG GAAGOCTACT TTGACCCCTC 60 GCTCCCGCAC ATGACGTACC GCGTGGTCGA GATTGTTGTT CTCTTCGTGC TTTCCTTTTG 120 sCTGATGGGT CAGTCTTCAC CCCTCGCGCT CGCTCTCGGC ATTGTCOTCA ocoscATCTC 180 TCAGGGTCGC TGCsGCTGGG TAATGCATGA GATGGGCCAT GGGTCGTTCA CTGGTGTCAT 240 TTGGCTTGAC sACCGGTTGT GCGAGTTCTT TTACGGCGTT GGTTsTOGCA TGAGCsGTCA 300 TTACTGGAAA AACCAGCACA GCAAACACCA CGCAGCGCCA AACCGGCTCG AGCACGATOT 360 AGATCTCAAC ACCTTGCCAT TGGTGGCCTT CAACGAOCGC GTCGTGCsCA AGOTCCGACC 420 (2) INFORMATION FOR SEQ ID NO: 20: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 125 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: not relevant (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: peptide (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 20 Arg Val Arg Pro Arg Val Arg Arg Glu Gln Leu He Lys Glu Gly 1 - 5 10 15 Tyr Phe Asp Pro Ser Leu Pro His Met Thr Tyr Arg Val Val Glu 20 25 30 He Val Val Leu Phe Val Leu Ser Phe Trp Leu Met Gly Gln Ser 35 40 45 Ser Pro Leu Ala Leu Ala Leu Gly He Val Val Ser Gly He Ser 50 55 60 Gln Gly Arg Cys Gly Trp Val Met His Glu Met Oly His Oly Ser 65 70 75 Phe Thr Gly Val He Trp Leu Asp Asp Arg Leu Cys Glu Phe Phe 65 70 75 Tyr Gly Val Gly Cys Gly Met Ser Gly His Tyr Trp Lys Asn Gln 80 85 90 His Ser Lys His His Wing Ala Pro Asn Arg Leu Glu His Asp Val 95 100 105 Asp Leu Asn Thr Leu Pro Leu Val Ala Phe Asn Glu Arg Val Val 110 115 120 Arg Lys Val Arg Pro 125 (2) INFORMATION FOR SEQ ID NO: 21: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1219 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (Contig Edita 2692004) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 21: GCACGCCGAC CGGCGCCGGG AGATCCTGGC AAAGTATCCA GAGATAAAGT CCTTGATGAA 60 ACCTGATCCC AATTTGATAT GGATTATAAT TATGATGGTT CTCACCCAGT TGGGTGCATT 120 TTACATAGTA AAAGACTTGG ACTGGAAATG GGTCATATTT GGGGCCTATG CGTTTGGCAG 180 TTGCATTAAC CACTCAATGA CTCTGGCTAT TCATGAGATT OCCCACAATG CTGCCTTTss 240 CAACTGCAAA GCAATGTGGA ATCGCTGGTT TGGAATGTTT GCTAATCTTC CTATTGGGAT 300 TCCATATTCA ATTTCCTTTA AGAGGTATCA CATGGATCAT CATCGGTACC TTGGAGCTGA 360 TGGCGTCGAT GTAGATATTC CTACCGATTT TGAGGGCTGG TTCTTCTGTA CCGCTTTCAG 420 AAAGTTTATA TGGGTTATTC TTCAOCCTCT CTTTTATsCC TTTCOACCTC TGTTCATCAA 480 CCCCAAACCA ATTACGTATC TGGAAGTTAT CAATACCGTG GCACAGGTCA CTTTTGACAT 540 TTTAATTTAT TACTTTTTGG GAATTAAATC CTTAGTCTAC ATsTTGGCAG CATCTTTACT 600 TGGCCTGGGT TTsCACCCAA TTTCTGGACA TTTTATAGCT OAOCATTACA TGTTCTTAAA 660 OGGTCATGAA ACTTACTCAT ATTATGGGCC TCTGAATTTA CTTACCTTCA ATGTGOGTTA 720 TCATAATGAA CATCATGATT TCCCCAACAT TCCTGGAAAA AGTCTTCCAC TGGTGAGGAA 780 AATAGCAGCT OAATACTATO ACAACCTCCC TCACTACAAT TCCTGOATAA AAGTACTGTA 840 TsATTTTGTG ATGGATGATA CAATAAGTCC CTACTCAAGA ATGAAsAOsC ACCAAAAAOG 900 AGAOATGGTG CTGGAGTAAA TATCATTAGT GCCAAAOOsA TTCTTCTCCA AAACTTTAGA 960 TGATAAAATG GAATTTTTGC ATTATTAAAC TTGAGACCAG TGATGCTCAG AAGCTCCCCT 1020 GGCACAATTT CAGAGTAAGA GCTCGGTGAT ACCAAGAAGT GAATCTGGCT TTTAAACAGT 1080 CAGCCTGACT CTGTACTGCT CAGTTTCACT CACAGGAAAC TTGTGACTTG TGTATTATCG 1140 TCATTGAGGA TGTTTCACTC ATGTCTGTCA TTTTATAAGC ATATCATTTA AAAAGCTTCT 1200 AAAAAGCTAT TTCGCCAGG 1219 (2) INFORMATION FOR SEQ ID NO: 22: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 655 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (Contig Edita 2153 * 526) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 22: TTACCTTCTA csTCcacTTC TTCCTCACTT ATGTGCCACT ATTOOOGCTG AAAGCTTCCT 60 GGGCCTTTTC TTCATAGTCA GGTTCCTGGA AAsCAACTGG TTTGTsTGGG TOACACAGAT 120 GAACCATATT CCCATGCACA TTGATCATGA CCGGAACATG GACTGGGTTT CCACCCAGCT 180 CCAGGCCACA TGCAATOTCC ACAAGTCTGC CTTCAATGAC tosttcAsts GACACCTCAA 240 CTTCCAGATT GAGCACCATC TTTTTCCCAC GATGCCTCGA CACAATTACC ACAAAGTGGC 300 TCCCCTGGTG CAGTCCTTGT GTGCCAAGCA TGGCATAGAG TACCAOTCCA AGCCCCTGCT 360 GTCAGCCTTC GCCGACATCA TCCACTCACT AAAGGAGTCA OGGCAGCTCT GGCTAGATsC 420 CTATCTTCAC CAATAACAAC AGCCACCCTG CCCAGTCTGG AAGAAGAGGA GGAAGACTCT 480 GOAsCCAAGG CAGAGGGGAG CTTGAGGGAC AATGCCACTA TAGTTTAATA CTCAGAGGGG 540 GTTGGGTTTG GGGACATAAA GCCTCTGACT CAAACTCCTC CCTTTTATCT TCTAGCCACA 600 GTTCTAAsAC CCAAAGTGGG GGGTGGACAC AGAAGTCCCT AGGAGsGAAG GAOCT 655 (2) INFORMATION FOR SEQ ID NO: 23: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 304 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (Cont.

Edited 3506132) (Í) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 23: GTCTTTTACT TTGGCAATGG CTGGATTCCT ACCCTCATCA COsCCTTTOT CCTTOCTACC 60 TCTCAGGCCC AAGCTGGATs GCTGCAACAT GATTATOGCC ACCTGTCTOT CTACAsAAAA 120 CCCAAGTGGA ACCACCTTGT CCACAAATTC GTCATTGGCC ACTTAAAGGG TGCCTCTOCC 180 AACTGGTGGA ATCATCGCCA CTTCCAGCAC CACOCCAAsC CTAACATCTT CCACAAOsAT 240 CCCGATGTGA ACATsCTGCA CGTGTTTGTT CTGGGCGAAT GCCAOCCCAT CGAOTACGCC 300 AAGA 304 (2) INFORMATION FOR SEQ ID NO: 24: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 918 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (Contig Edita 3854933) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 24: CAGGsACCTA CCCCGCGCTA CTTCACCTGG GACGAGGTGG CCCAGCGCTC AGGGTGCGAG 60 GAOCOOTOGC TAOTGATCGA CCGTAAGGTG TACAACATCA GCGAGTTCAC CCGCCGGCAT 120 CCAGGGGGCT CCCGGGTCAT CAGCCACTAC GCCGGGCAGG ATGCCACGGA TCCCTTTGTG 180 GCCTTCCACA TCAACAAGGG CCTTGTGAAG AAGTATATGA ACTCTCTCCT GATTGGAGAA 240 CTGTCTCCAO AGCAGCCCAG CTTTGAGCCC ACCAAGAATA AAGAGCTGAC AGATGAGTTC 300 CGGsAGCTGC GssCCACAGT GGAGCGGATG GGGCTCATGA AGGCCAACCA TGTCTTCTTC 360 CTGCTGTACC TGCTGCACAT CTTGCTGCTs GATOaTGCAG CCTGGCTCAC CCTTTGGGTC 420 TTTGOGACsT CCTTTTTGCC CTTCCTCCTC TGTGCGGTGC TGCTCACTGC AGTTCAGGCC 480 CAGGCTGOCT OGCTGCAGCA tsActttsos CACCTGTCGG TCTTCAGCAC CTCAAAGTGG 540 AACCATCTGC TACATCATTT TGTGATTGGC CACCTGAAGG GGGCCCCCGC CAGTTGGTGG 600 AACCACATGC ACTTCCAGCA CCATGCCAAG CCCAACTGCT TCCGCAAAGA CCCAGACATC 660 AACATOCATC CCTTCTTCTT TGCCTTGGGG AAGATCCTCT CTGTGGAGCT TGGGAAACAG 720 AAGAAAAAAT ATATGCCGTA CAACCACCAG CACARATACT TCTTCCTAAT TGGGCCCCCA 780 GCCTTGCTGC CTCTCTACTT CCAGTGGTAT ATTTTCTATT TTOTTATCCA GCGAAAGAAG 840 TGGGTGGACT TGGCCTGGAT CAGCAAACAG GAATACGATG AAGCCGGGCT TCCATTGTCC 918 900 ACCGCAAATG CTTCTAAA (2) INFORMATION FOR SEQ ID NO: 25: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1686 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (Contig Editad 2511785) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 25: GCCACTTAAA OGGTGCCTCT GCCAACTGOT GGAATCATCG CCACTTCCAG CACCACGCCA 60 AGCCTAACAT CTTCCACAAG GATCCCGATO TGAACATGCT GCACGTGTTT GTTCTGGGCG 120 AATGGCAGCC CATCGAGTAC GGCAAGAAGA AGCTGAAATA CCTGCCCTAC AATCACCAGC 180 ACGAATACTT CTTCCTGATT GGGCCGCCGC TGCTCATCCC CATGTATTTC CAGTACCAGA 240 TCATCATGAC CATGATCGTC CATAAGAACT GGGTGGACCT GGCCTGGGCC GTCAGCTACT 300 ACATCCGGTT CTTCATCACC TACATCCCTT TCTACGGCAT CCTGGGAGCC CTCCTTTTCC 360 TCAACTTCAT CAGGTTCCTG GAGAGCCACT GGTTTGTGTG GGTCACACAG ATGAATCACA 420 TCGTCATGGA GATTOACCAG GAOGCCTACC GTGACTGGTT CAGTAGCCAG CTGACAGCCA 480 CCTGCAACGT OGAOCAOTCC TTCTTCAACG ACTGGTTCAG TGGACACCTT AACTTCCAGA 540 TTGAGCACC? CCTCTTCCCC ACCATGCCCC GGCACAACTT ACACAAGATC GCCCCGCTGG 600 TGAAGTCTCT ATGTGCCAAs CATGGCATTG AATACCAGGA GAAGCCGCTA CTGAGGGCCC 660 TGCTGGACAT CATCAGGTCC CTGAAGAAGT CTGGGAAGCT GTGGCTGGAC GCCTACCTTC 720 ACAAATGAAG CCACAGCCCC CGGGACACCG TsGGGAAsGG GTGCAssTGG GsTGATGGCC 780 AGAGGAATGA TGGGCTTTTG TTCTGAsGGG TGTCCGAGAG GCTGGTsTAT GCACTGCTCA 840 CGGACCCCAT GTTGGATCTT TCTCCCTTTC TCCTCTCCTT TTTCTCTTCA CATCTCCCCC 900 ATAGCACCCT GCCCTCATGG GACCTGCCCT CCCTCAGCCG TCAGCCATCA GCCATOGCCC 960 TCCCAGTGCC TCCTAGCCCC TTCTTCCAAG GAGCAGAGAG sTGsCCACCG GGGGTGGCTC 1020 TGTCCTACCT CCACTCTCTG CCCCTAAAGA TGGGAGGAGA CCAGCGGTCC ATGGGTCTGG 1080 CCTGTGAOTC TCCCCTTOCA sCCTGGTCAC TAGGCATCAC CCCCGCTTTs GTTCTTCAGA 1140 TsCTCTTGGG GTTCATAGGG GCAGGTCCTA GTCGGGCAGG GCCCCTGACC CTCCCGGCCT 1200 GGCTTCACTC TCCCTGACGG CTGCCATTOG TCCACCCTTT CATAGAGAOG CCTsCTTTsT 1260 TACAAAGCTC GGGTCTCCCT CCTGCAGCTC GGTTAAGTAC CCGAGGCCTC TCTTAAGATG 1320 TCCAGGGCCC CAGGCCCGCG GGCACAGCCA GCCCAAACCT TGGGCCCTGG AAGAGTCCTC 1380 CACCCCATCA CTAGAGTGCT CTGACCCTGG GCTTTCACGG GCCCCATTCC ACCGCCTCCC 1440 CAACTTGAGC CTGTGACCTT GGGACCAAAG GGGGAGTCCC TCGTCTCTTG TGACTCAGCA 1500 GAGGCAsTGG CCACGTTCAG GGAGOOGCCG GCTGGCCTGG AGGCTCAGCC CACCCTCCAG 1560 CTTTTCCTCA GGGTGTCCTG AGOTCCAAAA TTCTGGAGCA ATCTGACCCT TCTCCAAAGG 1620 CTCTGTTATC AGCTsGGCAG TGCCAGCCAA TCCCTGGCCA TTTGGCCCCA GGGGACGTGG 1680 GCCCTG 1686 (2) INFORMATION FOR SEQ ID N0: 26: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1843 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY : linear (ii) TYPE OF MOLECULE: other nucleic acid (Contig 2535 (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 26: GTCTTTTACT TTGGCAATGG CTGGATTCCT ACCCTCATCA CGGCCTTTGT CCTTGCTACC 60 TCTCAGGCCC AAGCTGGATG GCTGCAACAT GATTATGGCC ACCTGTCTGT CTACAGAAAA 120 CCCAAGTGGA ACCACCTTGT CCACAAATTC GTCATTGGCC ACTTAAAGGG TGCCTCTGCC 180 AACTGGTGGA ATCATCGCCA CTTCCAGCAC CACGCCAAGC CTAACATCTT CCACAAGGAT 240 CCCGATGTGA ACATOCTOCA CGTGTTTsTT CTOGGCGAAT OOCAOCCCAT COAOTAOSGC 300 AAGAAGAAGC TGAAATACCT GCCCTACAAT CACCAGCACG AATACTTCTT CCTGATTGGG 360 CCGCCGCTGC TCATCCCCAT GTATTTCCAG TACCAGATCA TCATGACCAT GATCGTCCAT 420 AAGAACTGGG TGGACCTGGC CTGsGCCGTC AGCTACTACA TCCGGTTCTT CATCACCTAC 480 ATCCCTTTCT ACGGCATCCT GGGAGCCCTC CTTTTCCTCA ACTTCATCAG GTTCCTGGAG 540 AGCCACTGGT TTGTGTGGGT CACACAGATG AATCACATCG TCATGsAGAT TGACCAGGAG 600 GCCTACCGTG ACTGGTTCAG TAGCCAGCTG ACAGCCACCT GCAACGTGGA GCAGTCCTTC 660 TTCAACGACT sGTTCAGTGG ACACCTTAAC TTCCAGATTG AGCACCACCT CTTCCCCACC 720 ATGCCCCGGC ACAACTTACA CAAGATCGCC CCGCTGGTGA AGTCTCTATG TGCCAAGCAT 780 GGCATTGAAT ACCAGGAGAA GCCGCTACTG AGGGCCCTGC TGGACATCAT CAGGTCCCTG 840 AAGAAGTCTG GGAAGCTGTG GCTGGACGCC TACCTTCACA AATGAAGCCA CAGCCCCCGG 900 GACACCGTGG GGAAGGGGTG CAGGTGGGGT GATGGCCAGA GGAATGATGG GCTTTTGTTC 960 TGAGGGGTGT CCGAOAOsCT GGTGTATGCA CTsCTCACss ACCCCATOTT GGATCTTTCT 1020 CCCTTTCTCC TCTCCTTTTT CTCTTCACAT CTCCCCCATA OCACCCTOCC CTCATGGGAC 1080 CTGCCCTCCC.TCAGCCGTCA GCCATCAGCC ATGGCCCTCC CAGTOCCTCC TAOCCCCTTC 1140 TTCCAAGGAG CAGAGAGGTG GCCACCGGGG GTOGCTCTGT CCTACCTCCA CTCTCTGCCC 1200 CTAAAGATGG GAGGAGACCA GCGGTCCATG GGTCTGGCCT GTGAGTCTCC CCTTGCAGCC 1260 TGGTCACTAG GCATCACCCC CGCTTTGGTT CTTCAGATGC TCTTGGGGTT CATAGGGGCA 1320 GGTCCTAGTC GGGCAGGGCC CCTGACCCTC CCGGCCTGGC TTCACTCTCC CTGACGGCTG 1380 CCATTGGTCC ACCCTTTCAT AGAGAGGCCT GCTTTGTTAC AAAGCTCGGG TCTCCCTCCT 1440 GCAGCTCGGT TAAGTACCCG AGGCCTCTCT TAAGATGTCC AGGGCCCCAG GCCCGCGGGC 1500 ACAGCCAGCC CAAACCTTGG GCCCTGGAAG AGTCCTCCAC CCCATCACTA GÁGTGCTCTG 1560 ACCCTGGGCT TTCACGGGCC CCATTCCACC GCCTCCCCAA CTTGAGCCTG TGACCTTGGG 1620 ACCAAAGGGG GAGTCCCTCG TCTCTTGTGA CTCAGCAGAG GCAGTGGCCA CGTTCAGGGA 1680 GGGGCcsscT osccTssAOG CTCAGCCCAC CCTCCAGCTT TTCCTCAGGG TGTCCTGAGG 17 O TCCAAGATTC TGGAGCAATC TGACCCTTCT CCAAAGGCTC TGTTATCAGC TGGGCAsTGC 1800 CAGCCAATCC CTOsCCATTT OOCCCCAOGG GACGTGGsCC CTO 1843 (2) INFORMATION FOR SEQ ID NO: 27: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 2257 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: other nucleic acid (Contig Edited 253538a) (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 27: CAGGGACCTA CCCCGCGCTA CTTCACCTGG GACGAGGTGG CCCAGCGCTC AGGGTGCGAG 60 GAGCGGTGGC TAGTGATCGA CCGTAAssTG TACAACATCA sCGAGTTCAC CCGCCGGCAT 120 CCAGGGGOCT CCCOsOTCAT CAGCCACTAC GCCGOsCAGG ATGCCACOGA TCCCTTTsTG 180 GCCTTCCACA TCAACAAGGG CCTTGTGAAG AAGTATATGA ACTCTCTCCT GATTGGAGAA 2 0 CTGTCTCCAG AGCAGCCCAG CTTTGAGCCC ACCAAGAATA AAGAGCTGAC AGATGAGTTC 300 CGGGAGCTGC GGGCCACAGT GGAGCGGATG GGGCTCATGA AGGCCAACCA TGTCTTCTTC 360 CTGCTGTACC TGCTGCACAT CTTGCTGCTG GATGGTGCAG CCTGGCTCAC CCTTTGGGTC 420 TTTGGGACGT CCTTTTTTGCC CTTCCTCCTC TGTGCGGTGC TGCTCAGTGC AGTTCAGCAG 480 GCCCAAGCTG GATGGCTGCA ACATGATTAT GGCCACCTGT CTGTCTACAG AAAACCCAAG 540 TGGAACCACC TTGTCCACAA ATTCGTCATT GGCCACTTAA AGGGTGCCTC TGCCAACTGG 600 TGGAATCATC GCCACTTCCA GCACCACGCC AAGCCTAACA TCTTCCACAA GGATCCCGAT 660 GTGAACATGC TGCACGTGTT TGTTCTGGGC GAATGGCAGC CCATCGAGTA CGGCAAGAAG 720 AAGCTGAAAT ACCTGCCCTA CAATCACCAO CACGAATACT TCTTCCTGAT TGGGCCGCCG 780 CTGCTCATCC CCATGTATTT CCAGTACCAs ATCATCATGA CCATGATCGT CCATAAGAAC 840 TGGaTGGACC TGGCCTGGGC CGTCAGCTAC TACATCCGGT TCTTCATCAC CTACATCCCT 900 TTCTACGGCA TCCTGGGAGC CCTCCTTTTC CTCAACTTCA TCAGGTTCCT GGAGAGCCAC 960 TGsTTTGTGT GOGTCACAC? GATGAATCAC ATCGTCATGG AGATTGACCA GGAGGCCTAC 1020.

CGTGACTGGT TCAGTAGCCA GCTGACAGCC ACCTOCAACG TGGAGCAGTC CTTCTTCAAC 1080 GACTGGTTCA GTGGACACCT TAACTTCCAG ATTGAGCACC ACCTCTTCCC CACCATGCCC 1140 CssCACAACT TACACAAGAT COCCCCGCTG GTGAAGTCTC TATGTsCCAA sCATGGCATT 1200 GAATACCAGG AGAAGCCGCT ACTGAGGGCC CTGCTGGACA TCATCAGGTC CCTGAAGAAG 1260 TCTGGGAAsC TOTGGCTGGA CGCCTACCTT CACAAATGAA GCCACAGCCC CCGGGACACC 1320 GTGGGGAAGG GGTGCAGGTG GGGTGATGGC CAGAGGAATG ATGGGCTCTTT GTTCTGAGGG 1380 GTGTCCGAGA GGCTGGTGTA TGCACTGCTC ACGGACCCCA TGTTGGATCT TTCTCCCTTT 1440 CTCCTCTCCT TTTTCTCTTC ACATCTCCCC CATAGCACCC TGCCCTCATG GGACCTOCCC 1500 TCCCTCAGCC GTCAGCCATC AGCCATGGCC CTCCCAGTGC CTCCTAGCCC CTTCTTCCAA 1560 GsAsCAsAsA OOTOGCCACC GGGGGGTGsCT CTGTCCTACC TCCACTCTCT GCCCCTAAAG 1620 ATGGGAGGAG ACCAGCGGTC CATGGGTCTG GCCTGTGAOT CTCCCCTTGC AGCCTGGTCA 1680 CTAGGCATCA CCCCCGCTTT GGTTCTTCAG ATGCTCTTGG GGTTCATAGG GGCAGGTCCT 1740 AGTCGGsCAs GGCCCCTGAC CCTCCCGGCC TGGCTTCACT CTCCCTGACs GCTGCCATTO 1800 GTCCACCCTT TCATAGAGAG GCCTGCTTTG TTACAAAGCT CGGGTCTCCC TCCTGCAsCT 1860 CGGTTAAGTA CCCGAGGCCT CTCTTAAGAT GTCCAGGGCC CCAGGCCCGC GGGCACAsCC 1920 AGCCCAAACC TTGGGCCCTG GAAGAGTCCT CCACCCCATC ACTAGAGTGC TCTOACCCTO 1980 GGCTTTCACG GGCCCCATTC CACCGCCTCC CCAACTTGAG CCTGTOACCT TGGGACCAAA 2040 GGGGGAGTCC CTCGTCTCTT GTGACTCAGC AGAGGCAGTG GCCACGTTCA GGGAGGGGCC 2100 GGCTGGCCTG GAGGCTCAGC CCACCCTCCA GCTTTTCCTC AGGGTGTCCT GAGGTCCAAG 2160 ATTCTGGAGC AATCTGACCC TTCTCCAAAG GCTCTGTTAT CAGCTGGGCA GTGCCAGCCA 2220 ATCCCTGGCC ATTTGGCCCC AGGGGACGTG GGCCCTG 2257 (2) INFORMATION FOR SEQ ID NO: 28: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 411 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: amino acid (Translation of Cont 2692004) (XÍ) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 28: His Wing Asp Arg Arg Arg Glu He Leu Wing Lys Tyr Pro Olu He 1 5 10 15 Lys Ser Leu Met Lys Pro Asp Pro Asn Leu He Trp He He He 20 2S 30 Met Met Val Leu Thr Oln Leu sly Ala Phe Tyr He Val Lys Asp 35 40 45 Leu Asp Trp Lys Trp Val He Phe sly Ala Tyr Ala Phe sly Ser 50 55 60 Cys He Asn His Met Met Thr Leu Ala He His Olu He Ala His 65 70 75 Asn Ala Ala Phe sly Asn Cys Lys Ala Met Trp Asn Arg Trp Phe 80 85 90 Gly Met Phe Wing Asn Leu Pro He Gly He Pro Tyr Ser He Ser 95 100 105 Phe Lys Arg Tyr His Met Asp His His Arg Tyr Leu Oly Wing Asp 110 115 120 Gly Val Asp Val Asp He Pro Thr Asp Phe Glu Gly Trp Phe Phe 125 130 135 Cys Thr Wing Phe Arg Lys Phe He Trp Val He Leu Gln Pro Leu 140 145 150 Phe Tyr Ala Phe Arg Pro Leu Phe He Asn Pro Lys Pro He Thr 155 160 165 Tyr Leu Glu Val He Asn Thr Val Wing Gln Val Thr Phe Asp He 170 175 180 Leu He Tyr Tyr Phe Leu Gly He Lys Ser Leu Val Tyr Met Leu 185 190 195 Ala Ala Ser Leu Leu Gly Leu Gly Leu His Pro He Ser Gly His 200 205 210 Phe He Ala Siu His Tyr Met Phe Leu Lys Gly His Glu Thr Tyr 215 220 225 Ser Tyr Tyr Oly Pro Leu Asn Leu Leu Thr Phe Asn Val Oly Tyr 230 235 240 His Asn Glu His Kis Asp Phe Pro Asn He Pro Gly Lys Ser Leu 245 250 255 Pro Leu Val Arg Lys He Wing Wing Glu Tyr Tyr Asp Asn Leu Pro 260 265 270 His Tyr Asn Ser Trp He Lys Val Leu Tyr Asp Phe Val Met Asp 275 280 285 Asp Thr He Ser Pro Tyr Ser Arg Met Lys Arg His Gln Lys Gly 290 295 300 siu Met Val Leu Olu Xaa He Ser Leu Val Pro Lys Gly Phe Phe 305 310 315 Ser Lys Thr Leu Aßp Aßp Lys Met Glu Phe Leu His Tyr Xaa Thr 320 325 330 Xaa Asp Gln Xaa Cys Ser Glu Ala Pro Leu Ala Gln Phe Gln Ser 335 335 345 Lys Ser Ser Val He Pro Arg Ser Glu Ser Gly Phe Xaa Thr Val 350 355 360 Ser Leu Thr Leu Tyr Cys Ser Val Ser Leu Thr Oly Asn Leu Xaa 365 370 375 Leu Val Tyr Tyr Arg His Xaa Gly Cys Phe Thr His Val Cys His 380 385 390 Phe He Ser Be He Phe Lys Lys Leu Leu Lys Ser Tyr Phe Wing 400 405 410 Arg (2) INFORMATION FOR SEQ ID NO: 29: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 218 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Aminoside (Tradussión de Conti 2153526) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 29: Tyr Leu Leu Arg Pro Leu Leu Pro His Leu Cys Wing Thr He Gly 1 5 10 15 Wing Glu Being Phe Leu Gly Leu Phe Phe He Val Arg Phe Leu Glu 20 25 30 Being Asn Trp Phe Val Trp Val Thr Gln Met Asn His He Pro Met 35 40 45 His He Asp His Asp Arg Asn Met Asp Trp Val Ser Thr Gln Leu 50 55 60 Gln Wing Thr Cys Asn Val His Lys Ser Wing Phe Asn Asp Trp Phe 65 70 75 Ser Gly His Leu Asn Phe Gln He Olu His His Leu Phe Pro Thr 80 85 90 Met Pro Arg His Asn Tyr His Lys Val Ala Pro Leu Val Gln Ser 95 100 105 Leu Cys Ala Lys His Gly He Glu Tyr Gln Ser Lys Pro Leu Leu 110 115 120 Ser Ala Phe Ala Asp He He His Ser Leu Lys Glu Ser Oly Gln 125 130 135 Leu Trp Leu Asp Ala Tyr Leu His Gln Xaa Gln Gln Pro Pro Cys 140 145 150 Pro Val Trp Lys Lys Arg Arg Lys Thr Leu Glu Pro Arg Gln Arg 155 160 165 Gly Wing Xaa Gly Thr Met Pro Leu Xaa Phe Asn Thr Gln Arg Gly 170 175 180 Leu Gly Leu Gly Thr Xaa Ser Leu Xaa Leu Lys Leu Leu Pro Phe 185 190 195 He Phe Xaa Pro Gln Phe Xaa Asp Pro Lys Trp Gly Val Asp Thr 200 205 210 Olu Val Pro Arg Arg Glu Gly Wing 215 (2) INFORMATION FOR SEQ ID NO: 30: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 71 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: amino acid (Translation of Cont 3506132) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 30: Val Phe Tyr Phe Gly Asn Gly Trp He Pro Thr Leu He Thr Wing 1 5 10 15 Phe Val Leu Wing Thr Ser Gln Wing Gln Wing Gly Trp Leu Gln His 20 25 30 Asp Tyr Gly His Leu. Ser Val Tyr Arg Lys Pro Lys Trp Asn His 35 40 45 Leu Val His Lys Phe Val He Gly His Leu Lys Gly Ala Wing Wing 50 55 60 Asn Trp Trp Asn His Arg His Phe Gln His His Wing Lys Pro Asn 65 70 75 Leu Gly Glu Trp Gln Pro He Glu Tyr Gly Lys Xaa 80 85 (2) INFORMATION FOR SEQ ID NO: 31: (i) 'CHARACTERISTICS OF THE SEQUENCE:' (A) LENGTH: 306 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear ( ii) TYPE OF MOLECULE: amino acid (Translation of Cont 3854933) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 31: Gln Gly Pro Thr Pro Arg Tyr Phe Thr Trp Asp Glu Val Wing Gln 1 5 10 15 Arg Ser Gly Cys Glu Glu Arg Trp Leu Val He Asp Arg Lys Val 20 25 30 Tyr Asn He Ser Glu Phe Thr Arg Arg His Pro Gly Gly Ser Arg 35 40 45 Val He Ser His Tyr Wing Gly Gln Asp Wing Thr Asp Pro Phe Val 50 55 60 Wing Phe His He Asn Lys Oly Leu Val Lys Lys Tyr Met Asn Ser 65 70 75 Leu Leu He Gly Olu Leu Ser Pro Glu Gln Pro Ser Phe Glu Pro 80 85 90 Thr Lys Asn Lys Glu Leu Thr Asp Glu Phe Arg Glu Leu Arg Wing 95 100 105 Thr Val Glu Arg Met Gly Leu Met Lys Ala Asn His Val Phe Phe 110 115 120 Leu Leu Tyr Leu Leu His He Leu Leu Leu Asp Gly Wing Wing Trp 125 130 135 Leu Thr Leu Trp Val Phe Oly Thr Ser Phe Leu Pro Phe Leu Leu 140 145 150 Cys Wing Val Leu Leu Ser Wing Val Oln Wing Gln Wing Gly Trp Leu 155 160 165 Gln His Asp Phe Gly His Leu Ser Val Phe Ser Thr Ser Lys Trp 170 175 180 Asn His Leu Leu His His Phe Val He Gly His Leu Lys Gly Wing 185 190 195 Pro Wing Ser Trp Trp Asn His Met His Phe Gln His His Wing Lys 200 205 210 Pro Asn Cys Phe Arg Lys Asp Pro Asp He Asn Met His Pro Phe 215 220 225 Phe Phe Ala Leu Gly Lys He Leu Ser Val Glu Leu Gly Lys Gln 230 235 240 L ys Lys Lys Tyr Met Pro Tyr Asn His Oln His Xaa Tyr Phe Phe 245 250 255 Leu He Gly Pro Pro Wing Leu Leu Pro Leu Tyr Phe Gln Trp Tyr 260 265 270 He Phe Tyr Phe Val He Gln Arg Lys Lys Trp Val Asp Leu Wing 275 280 285 Trp He Ser Lys Gln siu Tyr Asp Glu Wing sly Leu Pro Leu Ser 290 295 300 Thr Wing Asn Wing Ser Lys 305 (2) INFORMATION FOR SEQ ID NO: 32: (i) CHARACTERISTICS OF THE SEQUENCE: ( A) LENGTH: 566 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: amino acid (Tradussion of Cont 2511785) (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 32: His Leu Lys Gly Wing Ser Wing Asn Trp Trp Asn His Arg His Phe 1 5 10 15 Gln His His Ala Lys Pro Asn He Phe His Lys Asp Pro Asp Val 20 25 30 Asn Met Leu His Val Phe Val Leu Gly Glu Trp Gln Pro He Glu 35 40 45 Tyr Gly Lys Lys Lys Leu Lys Tyr Leu Pro Tyr Asn His Gln His 50 55 60 Glu Tyr Phe Phe Leu He Gly Pro Pro Leu Leu He Pro Met Tyr 65 70 75 Phe Oln Tyr Gln He He Met Met Thr Met He Val His Lys Asn Trp 80 85 90 Val Asp Leu Wing Trp Wing Val Ser Tyr Tyr He Arg Phe Phe He 95 100 105 Thr Tyr He Pro Phe Tyr Gly He Leu Gly Ala Leu Leu Phe Leu 110 115 120 Asn Phe He Arg Phe Leu Glu Ser His Trp Phe Val Trp Val Thr 125 130 135 Gln Met Asn His He Val Met Glu He Asp Gln Glu Ala Tyr Arg 140 145 150 Asp Trp Phe Be Ser Gln Leu Thr Ala Thr Cys Aßn Val Glu Gln 155 160 165 Being Phe Phe Asn Asp Trp Phe Ser Gly His Leu Asn Phe Gln He 170 175 180 Glu His His Leu Phe Pro Thr Met Pro Arg His Asn Leu His Lys 185 190 195 He Ala Pro Leu Val Lys Ser Leu Cys Ala Lys His Gly He Glu 200 205 210 Tyr Gln Glu Lys Pro Leu Leu Arg Wing Leu Leu Asp He He Arg 215 220 225 Ser Leu Lys Lys Ser Gly Lys Leu Trp Leu Asp Wing Tyr Leu His 230 235 240 Lys Xaa Ser His Ser Pro Arg Asp Thr Val Gly Lys Gly Cys Arg 245 250 255 Trp Gly Asp Gly Gln Arg Asn Asp Gly Leu Leu Phe Xaa Gly Val 260 265 270 Ser Glu Arg Leu Val Tyr Ala Leu Leu Thr Asp Pro Met Leu Asp 275 280 285 Leu Ser Pro Phe Leu Leu Ser Phe Phe Ser Ser His Leu Pro His 290 295 300 Ser Thr Leu Pro Ser Trp Asp Leu Pro Ser Leu Ser Arg Gln Pro 305 310 315 Be Wing Met Wing Leu Pro Val Pro Pro Be Pro Phe Phe Gln Gly 320 325 330 Wing Glu Arg Trp Pro Pro Gly Val Wing Leu Ser Tyr Leu His Ser 335 340 345 Leu Pro Leu Lys Met Gly Gly Asp Gln Arg Ser Met Gly Leu Ala 350 355 360 Cys Glu Ser Pro Leu Wing Wing Trp Ser Leu Gly He Thr Pro Wing 365 370 375 Leu Val Leu Gln Met Leu Leu Gly Phe He Gly Wing Gly Pro Ser 380 385 390 Arg Ala Gly Pro Leu Thr Leu Pro Wing Trp Leu His Ser Pro Xaa 400 405 410 Arg Leu Pro Leu Val His Pro Phe He Glu Arg Pro Wing Leu Leu 415 420 425 Gln Ser Ser Gly Leu Pro Pro Ala Wing Arg Leu Ser Thr Arg Gly 430 435 440 Leu Ser Xaa Aßp Val Oln Gly Pro Arg Pro Wing Gly Thr Wing Ser 445 450 455 Pro Asn Leu Gly Pro Trp Lys Ser Pro Pro Pro His His Xaa Ser 460 465 470 Wing Leu Thr Leu Gly Phe His Gly Pro His Be Thr Ala Ser Pro 475 480 485 Thr Xaa Wing Cys Asp Leu sly Thr Lys Gly Val Pro Arg Leu 490 495 500 Leu Xaa Leu Ser Arg Oly Ser Gly His Val Gln Gly Gly Wing Gly 505 510 515 Trp Pro Gly Gly Ser Wing His Pro Pro Wing Phe Pro Gln Gly Val 520 525 530 Leu Arg Ser Lys He Leu Olu Gln Ser Asp Pro Ser Lys Wing 535 540 545 Leu Leu Ser Wing Gly Gln Cys Gln Pro He Pro Gly His Leu Wing 550 555 560 Pro Gly Asp Val Oly Pro Xaa 565 (2) INFORMATION FOR SEQ ID NO: 33: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 619 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: amino acid (Translation of Cont 2535) (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 33: Val Phe Tyr Phe Gly Asn Gly Trp lie Pro Thr Leu He Thr Wing 1 5 10 15 Phe Val Leu Wing Thr Ser Gln Wing Gln Wing Gly Trp Leu Gln His 20 25 30 Asp Tyr Oly His Leu Ser Val Tyr Arg Lys Pro Lys Trp Asn Hiß 35 40 45 Leu Val His Lys Phe Val He Gly His Leu Lys Gly Ala Ser Wing 50 55 60 Asn Trp Trp Asn His Arg His Phe Gln His His Ala Lyß Pro Aßn 65"70 75 He Phe His Lys Asp Pro Asp Val Asn Met Leu His Val Phe Val 80 85 90 Leu Gly Glu Trp Gln Pro He Glu Tyr Oly Lys Lys Lys Leu Lys 95 100 105 Tyr Leu Pro Tyr Asn His Gln His Olu Tyr Phe Phe Leu He Gly 110 115 120 Pro Pro Leu Leu He Pro Met Tyr Phe Oln Tyr Gln He He Met Met 125 130 135 Thr Met He Val His Lys Asn Trp Val Aßp Leu Wing Trp Wing Val 140 145 150 Ser Tyr Tyr He Arg Phe He Thr Tyr He Pro Phe Tyr Gly 155 160 165 He Leu Gly Ala Leu Leu Phe Leu Asn Phe He Arg Phe Leu Glu 170 175 180 Ser His Trp Phe Val Trp Val Thr Gln Met Asn His He Val Val Met 185 190 195 Glu He Asp Gln Glu Ala Tyr Arg Asp Trp Phe Ser Ser Gln Leu - 200 205 210 Thr Ala Thr Cys Asn Val Glu Gln Ser Phe Phe Asn Asp Trp Phe 215 220 225 Ser Oly His Leu Asn Phe Gln He Glu His His Leu Phe Pro Thr 230 235 240 Met Pro Arg Hiß Asn Leu His Lys He Ala Pro Leu Val Lys Ser 245 250 255 Leu Cys Ala Lys His Gly He Glu Tyr Gln Glu Lys Pro Leu Leu 260 265 270 Arg Ala Leu Leu Asp He He Arg Ser Leu Lys Ser Gly Lys 275 280 285 Leu Trp Leu Asp Ala Tyr Leu His Lys Xaa Ser His Ser Pro Arg 290 295 300 Asp Thr Val Gly Lys Gly Cys Arg Trp Gly Asp Gly Gln Arg Asn 305 310 315 Asp Gly Leu Leu Phe Xaa Gly Val Ser Glu Arg Leu Val Tyr Ala 320 325 330 Leu Leu Thr Asp Pro Met Leu Asp Leu Ser Pro Phe Leu Leu Ser 335 340 345 Phe Phe Ser Be His Leu Pro His Ser Thr Leu Pro Ser Trp Asp 350 355 360 Leu Pro Ser Leu Ser Arg Gln Pro Ser Ala Ala Ala Leu Pro Val 3 , 65 370 375 Pro Pro Ser Phe Pro Phe Gln sly Ala Glu Arg Trp Pro Pro sly 380 385 390 Val Ala Leu Ser Tyr Leu His Ser Leu Pro Leu Lys Met Gly Gly 400 405 410 Asp Gln Arg Ser Met Gly Leu Ala Cys Glu Ser Pro Leu Ala Ala 415 420 425 Trp Ser Leu Gly He Thr Pro Wing Leu Val Leu Gln Met Leu Leu 430 435 440 Gly Phe He Gly Wing Gly Pro Ser Arg Wing Gly Pro Leu Thr Leu 445 450 455 Pro Ala Trp Leu His Ser Pro Xaa Arg Leu Pro Leu Val His Pro 460 465 470 Phe He Glu Arg Pro Ala Leu Leu Gln Ser Ser Gly Leu Pro Pro 475 480 485 Wing Wing Arg Leu Ser Thr Arg Gly Leu Ser Xaa Asp Val Gln sly 490 495 500 Pro Arg Pro Wing Gly Thr Wing Pro Pro Asn Leu Gly Pro Trp Lys 505 510 515 Ser Pro Pro Pro His His Xaa Ser Ala Leu Thr Leu Gly Phe His 520 525 530 Gly Pro His Ser Thr Ala Ser Pro Thr Xaa Wing Cys Asp Leu Oly 535 540 545 Thr Lys sly Gly Val Pro Arg Leu Leu Xaa Leu Ser Arg Gly Ser 550 555 560 Gly His Val Gln Gly Gly Wing sly Trp Pro Gly Gly Ser Wing His 565 570 575 Pro Pro Wing Phe Pro Gln siy Val Leu Arg Ser Lys He Leu Olu 580 585 590 Oln Ser Asp Pro Pro Lyß Ala Leu Leu Ser Ala Gly Oln Cys 595 600 605 Oln Pro He Pro Gly His Leu Ala Pro Gly Asp Val sly Pro Xaa 610 615 620 (2 ) INFORMATION FOR SEQ ID NO: 34: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 757 amino acids (B) TYPE: amino acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (i) i) TYPE OF MOLECULE: amino acid (Tradussión de Conti 253538a) (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 34: Gln Gly Pro Thr Pro Arg Tyr Phe Thr Trp Asp Glu Val Wing Gln 1 5 10 15 Arg Ser Gly Cys Glu Glu Arg Trp Leu Val He Asp Arg Lys Val 20 25 30 Tyr Asn He Ser Siu Phe Thr Arg Arg His Pro Gly Gly Ser Arg 35 40 45 Val He Ser His Tyr Wing Gly Gln Asp Wing Thr Asp Pro Phe Val 50 55 60 Wing Phe His He Asn Lys Gly Leu Val Lys Lys Tyr Met Asn Ser 65 70 75 Leu Leu He Oly Glu Leu Ser Pro Glu Gln Pro Ser Phe Glu Pro 80 85 90 Thr Lys Asn Lys Glu Leu Thr Asp Glu Phe Arg Glu Leu Arg Wing 95 100 105 Thr Val Glu Arg Met Gly Leu Met Lys Ala Asn His Val Phe Phe 110 115 120 Leu Leu Tyr Leu Leu His He Leu Leu Leu Asp Gly Ala Wing Trp 125 130 135 Leu Thr Leu Trp Val Phe Gly Thr Ser Phe Leu Pro Phe Leu Leu 140 145 150 Cys Ala Val Leu Leu Ser Ala Val Gln Gln Ala Oln Ala Oly Trp 155 160 165 Leu Gln Hiß Asp Tyr Gly His Leu Ser Val Tyr Arg Lys Pro Lys 170 175 180 Trp Asn His Leu Val His Lys Phe Val He Oly His Leu Lys Gly 185 190 195 Wing Ser Wing Asn Trp Trp Asn His Arg His Phe Gln His His Wing 200 205 210 Lys Pro Asn He Phe His Lys Asp Pro Asp Val Asn Met Leu His 215 220 225 Val Phe Val Leu Gly Glu Trp Gln Pro He Glu Tyr Gly Lys Lys 230 235 240 Lys Leu Lys Tyr Leu Pro Tyr Asn His Gln His Olu Tyr Phe Phe 245 250 255 Leu He Gly Pro Pro Leu Leu He Pro Met Tyr Phe Gln Tyr Gln 260 265 270 He He Met Met Thr Met He Val His Lys Asn Trp Val Asp Leu Ala 275 280 285 Trp Wing Val Ser Tyr Tyr He Arg Phe Phe He Thr Tyr He Pro 290 295 300 Phe Tyr Gly He Leu Gly Ala Leu Leu Phe Leu Asn Phe He Arg 305 310 315 Phe Leu Glu Ser His Trp Phe Val Trp Val Thr Gln Met Asn His 320 325 330 He Val Met Glu He Aßp Gln Glu Ala Tyr Arg Asp Trp Phe Ser 335 340 345 Ser Gln Leu Thr Ala Thr Cys Asn Val Glu Gln Ser Phe Phe Aßn 350 355 360 Asp Trp Phe Ser Oly His Leu Asn Phe Gln He Glu His His Leu 365 370 375 Phe Pro Thr Met Pro Arg His Asp Leu His Lys He Wing Pro Leu 380 385 390 Val Lys Ser Leu Cys Wing Lys His Gly He Glu Tyr Gln Glu Lys 400 405 410 Pro Leu Leu Arg Ala Leu Leu Asp He He Arg Ser Leu Lys Lys 415 420 425 Ser Gly Lys Leu Trp Leu Asp Wing Tyr Leu His Lys Xaa Ser His 430 435 440 Ser Pro Arg Asp Thr Val Gly Lys Gly Cys Arg Trp Gly Asp Gly 445 450 455 Gln Arg Asn Asp Oly Leu Leu Phe Xaa Gly Val Ser Olu Arg Leu 460 465 470 Val Tyr Ala Leu Leu Thr Asp Pro Met Leu Asp Leu Ser Pro Phe 475 480 485 Leu Leu Ser Phe Phe Ser Ser Leu Pro His Ser Thr Leu Pro 490 495 500 Ser Trp Asp Leu Pro Ser Leu Ser Arg Gln Pro Ser Ala Ala Ala 505 510 515 Leu Pro Val Pro Pro Ser Pro Phe Phe Gln Gly Wing Glu Arg Trp 520 525 530 Pro Pro Gly Val Ala Leu Ser Tyr Leu His Ser Leu Pro Leu Lys 535 540 545 Met Gly Gly Asp Gln Arg Ser Met Gly Leu Ala Cys Glu Ser Pro 550 555 560 Leu Ala Ala Trp Ser Leu Gly He Thr Pro Ala Leu Val Leu Gln 565 570 575 Met Leu Leu Gly Phe He Gly Wing Gly Pro Ser Arg Wing Gly Pro 580 585 590 Leu Thr Leu Pro Wing Trp Leu His Ser Pro Xaa Arg Leu Pro Leu 595 600 605 Val His Pro Phe He Glu Arg Pro Wing Leu Leu Gln Ser Ser Gly 610 615 620 Leu Pro Pro Ala Ala Arg Leu Ser Thr Arg Gly Leu Ser Xaa Asp 625 630 635 Val Gln Gly Pro Arg Pro Wing Gly Thr Wing Pro Pro Asn Leu Gly 640 645 650 Pro Trp Lys Ser Pro Pro Pro His His Xaa Ser Ala Leu Thr Leu 655 660 665 Gly Phe His Gly Pro His Ser Thr Wing Ser Pro Thr Xaa Wing Cys 670 675 680 Asp Leu Gly Thr Lys Gly Oly Val Pro Arg Leu Leu Xaa Leu Ser 685 690 695 Arg Gly Ser Gly His Val Gln Gly Gly Wing Gly Trp Pro Gly Gly 700 705 710 Ser Ala His Pro Pro Wing Phe Pro Gln Gly Val Leu Arg Ser Lyß 715 720 725 He Leu Glu Gln Ser Asp Pro Ser Pro Lys Ala Leu Leu Ser Wing 730 735 740 Gly Gln Cys Gln Pro He Pro Gly His Leu Wing Pro Gly Asp Val 745 750 755 Gly Pro Xaa

Claims (88)

1. NOVELTY OF THE INVENTION Having described the foregoing invention, it is considered as a novelty and, therefore, the property contained in the following CLAIMS is resounded as such: 1. An isolated nucleic acid that appears: a nucleotide sesuensia represented in a SEQ ID NO. N0.-1.
2. 2. A polypeptide encoded by the nucleo acid of claim 1.
3. 3. A purified or isolated polypeptide comprising an amino acid sequence depicted in SEQ ID NO: 2.
4. 4. An isolated solid nucleic which sodifills the polypeptide of SEQ ID NO:
5. 5. A nucleic acid comprising: a nucleotide sequence that sodes a polypeptide that desaturates a fatty acid molecule at the carbon 5 of the carboxyl end of the fatty acid molecule.
6. 6. The isolated nucleic acid according to claim 5, characterized in that the nucleotide sequence is derived from a eukaryotic cell.
7. 7. The isolated nucleic acid according to claim 6, characterized in that the eukaryotic cell is a fungal cell.
8. 8. The isolated nucleic acid according to claim 7, characterized in that the fungus cell is of the genus Mortierella.
9. 9. The nusleiso isolate of sonformity are the reslamado in the reivindisasión 8, sarasterizado because the sélüla of Mortierella is of the espesie Mortierella alpina.
10. 10. The isolated nusleiso isolated from sonicity are the reslamado in claim 5, which is sarasterized because the sessile of nusleotides is fixed to a sesuensia of nusleotides represented in SEQ ID N0: 1.
11. 11. The nucleic acid isolated according to claim 10, characterized in that the sequence of nucleotides sodifices a sessensia of amino acids represented in SEQ ID NO: 2.
12. 12. The isolated nusleiso isolated from sonification are the reslamation in the claim 11, sarasterized because the amino acid sessence represented in SEQ ID NO: 2 is selessiona between the group that was in the amino acid residues 30-38, 41-44, 171-175, 203-212, and 387-394.
13. 13. An isolated or purified polypeptide that desaturates a fatty acid molecule in the sarbox of the sarboxyl end of the fatty acid molecule.
14. 14. A single isolated nusleiso that suffers from a nusleotide sesuensia that is substantially identical to a sesuensia of minus 50 nusleotides in SEQ ID NO: l.
15. 15. An isolated nucleic acid sequence having less than about 50% identity is SEQ ID NO: 1.
16. 16. A nucleic acid construct comprising: a nucleotide sequence depicted in SEQ ID NO: 1 linked to a heterologous nucleic acid.
17. 17. A nucleic acid construct comprising: a nucleotide sequence represented in a SEQ ID N0: 1 operably linked to a promoter.
18. 18. The isolated nucleic acid construct according to claim 17, characterized in that the promoter is functional in a microbial cell.
19. 19. The isolated nucleic acid construct according to claim 18, characterized in that the microbial cell is a yeast cell.
20. 20. The isolated nucleic acid construct according to claim 17, characterized in that the sequencing of nucleotides is derived from a fungus.
21. 21. The nucleic acid according to claim 19, characterized in that the fungus is of the genus Mortierella.
22. 22. The nucleic acid according to claim 20, characterized in that the fungus is from the Mortierella alpina thickene.
23. 23. A nucleic acid construct comprising: a sequencing of nusleotides that sodifies a polypeptide comprising an amino acid sequence that corresponds or is complementary to an amino acid sequence represented in SEQ ID NO: 2, wherein the nucleotide sesuensia is operably linked to a promoter that is functional in a host cell, wherein the nucleotide sequence encodes a polypeptide that desaturates a fatty acid molecule at the carboxyl end carbon 5 of a fatty acid molecule.
24. 24. A nucleic acid construct that somersates: a nucleotide sequence that encodes a functionally active? 5-desaturase, the desaturase having an amino acid sequence that corresponds or is complementary to all or a portion of the amino acid sequence depicted in SEQ ID. N0: 2, wherein the nucleotide sequence is operably linked to a functional promoter in a host cell.
25. 25. A recombinant yeast cell comprising: a nucleic acid construct according to claim 23 or claim 24.
26. 26. The recombinant yeast cell according to claim 25, characterized in that the cell of Yeast is a Saccharomyces cell.
27. 27. A host cell comprising: at least one copy of a nucleotide sequence encoding a polypeptide that converts the dihomo-γ-linolenic acid into arachidonic acid, wherein the misrobial cell or an ancestor of that microbial cell was transformed with a vector comprising that nucleotide sequence, and characterized in that the sequencing of nusleotides is operably linked to a funsional promoter in the host cell.
28. 28. The misrobial cell of soundness with that claimed in claim 27, sarasterized because the cell is a selented host cell of the group that was part of a fungal cell and an algae cell.
29. 29. The microbial cell according to claim 28, characterized in that the fungal cell is sucking minus one yeast cell and the algae cell is a seaweed cell.
30. 30. The compliant misrobial cell is the reslamation in claim 27, characterized in that the cell is enriched by 20: 3 fatty acids in sompasion are the host cell that lacks that sessile of nusleotides.
31. 31. The misrobial cell of soundness is the reslamation in claim 27, which is sarasterized because the cell is enriched by 20: 4 or? -3: 20: 4 fatty acids in somparasion are a host cell that lacks DNA sesuensia.
32. 32. The misrobial cell of sonformity is claimed in claim 27, characterized in that the cell is enriched by 20: 5 fatty acids compared to a host cell lacking the DNA sequence.
33. 33. The microbial cell as claimed in claim 27, characterized in that the cell has an altered amount of 20: 3 (8, 11, 14) fatty acids compared to a non-transformed microbial cell.
34. 34. A method for the production of an arachidonic acid in a culture of microbial cells, the method comprising: cultivating a culture of microbial cells having a plurality of microbial cells, wherein the microbial cells or their ancestors were transformed with a vector that comprises one or more nucleic acids having a nucleotide sequence that encodes a polypeptide that converts dihomo-7-linolenic acid into arachidonic acid, wherein the one or more nucleic acids are operably linked to a promoter, under conditions wherein the one or more nucleic acids are expressed and arachidonic acid is produced in the microbial cell culture.
35. 35. The method according to the claim in claim 34, sarasterized because the polypeptide is an enzyme that desaturates a molecule of fatty acids at the carbon 5 of the carboxyl end of the fatty acid molecule.
36. 36. The method of conformance with that claimed in claim 34, characterized in that the nucleotide sequence is derived from a Mortierella species.
37. 37. The method according to claim 37, characterized in that the dihomo-7-linolenic acid is supplied exogenously.
38. 38. The method of conformance with what is recited in claim 34, characterized in that the microbial cells are yeast cells.
39. 39. The method according to claim 38, characterized in that the yeast cells are cells of Saccharomyces species.
40. 40. The method according to claim 34, characterized in that the conditions are inducible.
41. 41. A resomminating yeast cell that converts more than about 5% of a 20: 3 fatty acid into a 20: 4 fatty acid.
42. 42. A nucleic acid probe comprising: a nucleotide sequence as represented by SEQ ID NO: 1
43. 43. A somatic host cell: a nucleic acid construct as claimed in claim 23 or claim 24
44. 44. A somatic host cell: a vestige including a nucleic acid encoding a fatty acid desaturase derived from Mortierella alpina, wherein the fatty acid desaturase comprises an amino acid sequence represented by SEQ ID NO: 2, wherein the nucleic acid is operably linked to a promoter.
45. 45. The host cell according to claim 44, characterized in that the host cell is a eukaryotic cell.
46. 46. The host cell according to claim * in claim 45, sarasterized because that eukaryotic cell is separated from the group that was in a mammalian cell, a plant cell, a fungal cell, a bird cell and a cell of seaweed
47. 47. The host cell according to claim claimed in claim 45 characterized in that the host cell contains dihomo-α-linolenide acid.
48. 48. The host cell according to the recitation in claim 45, characterized in that the host cell contains eicosapentaenoisoic acid.
49. 49. The host cell according to claim 44, characterized in that the promoter is delivered exogenously.
50. 50. A method for desaturating a dihomo-α-linolenic acid, the method comprising: culturing a recombinant microbial cell according to claim 37, under conditions suitable for the expression of the polypeptide encoded by the nucleic acid, characterized in that the The host cell further comprises a fatty acid substrate of said polypeptide.
51. 51. A fatty acid desaturated by the method according to claim 50.
52. 52. An oil comprising a fatty acid according to claim 51.
53. 53. A method for obtaining the biosynthesis of poly fatty acids. - altered long chain unsaturates comprising the steps of: cultivating a microbe that has cells that contain a transgene that encodes a transgene expression product which desaturates a molecule of fatty acids at the carbon 5 of the carboxyl end of the acid molecule fatty acids, characterized in that the transgene is operably associated with an expression control sequence, under condi tions by means of which the transgene is expressed, whereby the biosynthesis of long chain polyunsaturated fatty acids in said cells is altered.
54. 54. A method for obtaining the biosynthesis of altered long-chain polyunsaturated fatty acids comprising the steps of: cultivating a microbe that has cells containing a transgene, derived from a fungus or alga, encoding a transgene expression product which desaturates a fatty acid molecule at the carbon 5 of the carboxyl end of the fatty acid molecule, characterized in that the transgene is operably asosia is a sesuensia of expression control, under conditions whereby the one or more transgenes are expressed, whereby the biosynthesis of long chain polyunsaturated fatty acids in said cells is altered.
55. 55. The method according to claim 53 or 54, characterized in that the long chain polyunsaturated fatty acid is selected from the group consisting of arachidonic acid, dihomo-gamma-linolenic acid and eicosapentaenoic acid.
56. 56. A microbial oil or fraction thereof produced in accordance with the method of claim 53 or 54.
57. 57. A method for treating or preventing malnutrition comprising administering the microbial oil in accordance with what is claimed in claim 56 to a parent. there is a need for said treatment or prevention in an amount sufficient to effect the treatment or prevention.
58. 58. A pharmaceutical composition comprising the microbial oil or fraction according to claim 54 and a pharmaceutically acceptable carrier.
59. 59. The pharmaceutical composition according to claim 58, characterized in that the pharmaceutical composition is in the form of a solid or a liquid.
60. 60. The pharmaceutical composition according to claim claimed in claim 59, characterized in that the pharmaceutical composition is in a capsule or in the form of a tablet.
61. 61. The pharmaceutical composition according to claim 58, further comprising at least one nutrient selected from the group consisting of a vitamin, a mineral, a carbohydrate, a sugar, an amino acid, a free fatty acid, a phospholipid , an anti-oxidant, and a phenolic compound.
62. 62. A nutritional formula comprising the microbial oil or fraction thereof of claim 56.
63. 63. The nutritional formula according to claim 22, characterized in that the nutritional formula is selected from the group consisting of a formula for infants, a dietary supplement, and a dietary substitute.
64. 64. The nutritional formula of conformity is claimed in claim 63, which is sarasterized because the formula for infants, the dietary supplement or the dietetic substitute is in the form of liquid or solid.
65. 65. A formula for the presence of the microbial oil or fraction thereof according to claim 56.
66. The infant formula according to claim 1, further comprising at least one macronutrient sequestered from the group that were in coconut oil, soybean oil, cañola oil, mono and diglycerides, glusosa, edible lactose, electrodialyzed serum, electrialized skimmed leshe, leshe serum, soy protein, and other protein hydrolysates.
67. 67. The infant formula according to claim 66, further comprising at least one vitamin selected from the group consisting of Vitamins A, C, D, E, and B complex; and at least one mineral selected from the group consisting of calcium, magnesium, zinc, manganese, sodium, potassium, phosphorus, copper, chlorine, iodine, selenium, and iron.
68. 68. A dietary supplement comprising the microbial composition or fraction thereof according to the recitation in claim 56.
69. 69. The compliance dietary supplement is claimed in claim 68 which further comprises at least one macronutrient selected from the group consisting of in coconut oil, soybean oil, canola oil, mono and diglycerides, glucose, edible lactose, electrodialyzed whey, electrodialyzed skim milk, whey, soy protein, and other protein hydrolysates.
70. 70. The dietary supplement of sonformity is claimed in claim 69 which also comprises at least one vitamin selessionada of the group that you were in Vitamins A, C, D, E and B B-complex; and suando less a selessionado mineral of the group that you were in salsio, magnesium, zins, manganese, sodium, potassium, phosphorus, envelope, sloro, iodine, selenium, and iron.
71. 71. The dietidiosis supplement of soundness is the reslamation in claim 68 or claim 70, which is sarasterized because the dietary supplement is administered to a human or an animal.
72. 72. A dietetic substitute comprising the microbial oil or frassión of the same of sonformity is the reslamado in the reivindisasión 56.
73. 73. The dietetic substitute of sonformidad are the reslamado in the reivindisasión 72 that also somprende suando less a macronutriente selected from the group that is in bland soup, soy sauce, sauces, soy sauce, mono and diglycerides, glusosa, somestible poultice, whey elestrodialized, less dessremada elestrodializada, leshe serum, soy protein, and other protein hydrolysates.
74. 74. The dietitian substitute of sonformity is the reslamado in the vindication 73 that also sucks less sula a selessionada vitamin of the group that you were in Vitamins A, C, D, E and B somplejo; and suando less a selessionado mineral of the group that you were in salsio, magnesium, zins, manganese, sodium, potassium, phosphorus,, chlorine, iodine, selenium, and iron.
75. 75. The dietetic substitute according to claim claimed in claim 72 or claim 74, characterized in that the dietary substitute is administered to a human or an animal.
76. 76. A method for treating a patient having a condition caused by ingestion or insufficient production of polyunsaturated fatty acids comprising administering to the patient the dietary substitute of claim 72 or the dietary supplement of claim 68 in an amount sufficient to effect the treatment.
77. 77. The method according to claim 22, characterized in that the dietary substitute or the dietary supplement is administered enterally or parenterally.
78. 78. A cosmetic comprising the microbial oil or fraction thereof in accordance with that claimed in claim 56.
79. 79. The cosmetic according to claim claimed in claim 78, characterized in that the sosmetic is applied topically.
80. 80. The pharmaceutical composition according to claim 58, characterized in that the pharmaceutical composition is administered to a human or an animal.
81. 81. An animal feed that suffers the misrobial aseptic or the frassión of the same of sonformity are the reslamado in the reivindicación 56.
82. 82. The method of conformity with the claimed in claim 54 sarasterizado because the fungus is Mortierella spesies.
83. 83. The sonicity method is the reslamado in the vindication 82 where the fungus is Mortierella alpina.
84. 84. An isolated sessile organism sessile of the group that was in SEQ ID NO: 13 and SEQ ID NO: 15.
85. 85. A nusleotide sessence isolated from the group that was in SEQ ID NO: 7 and SEQ ID NO: 19.
86. 86 A sesuensia of isolated nusleotides that suffers from a sessile syndrome of selenced nusleotides of the group that was in SEQ ID NO: 13; SEQ ID NO: 15; SEQ ID NO: 17; SEQ ID NO: 19; SEQ ID NO: 21; SEQ ID NO: 22; SEQ ID NO: 23; SEQ ID NO: 24; SEQ ID NO: 25; SEQ ID NO: 26 and SEQ ID NO: 27.
87. 87. An isolated peptide sessence that results in a peptide sesession selessioned from the group that was in SEQ ID NO: 14; SEQ ID NO: 16; SEQ ID NO: 18; SEQ ID NO: 20; SEQ ID NO: 28; SEQ ID NO: 29; SEQ ID NO: 30; SEQ ID NO: 31; SEQ ID NO: 32; SEQ ID NO: 33 and SEQ ID NO: 34.
88. 88. Purified polypeptides produced from the nusleotide sesuensia of claims 84-86.