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US20080141390A1 - Plant genes involved in nitrate uptake and metabolism - Google Patents

Plant genes involved in nitrate uptake and metabolism Download PDF

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Publication number
US20080141390A1
US20080141390A1 US11/876,534 US87653407A US2008141390A1 US 20080141390 A1 US20080141390 A1 US 20080141390A1 US 87653407 A US87653407 A US 87653407A US 2008141390 A1 US2008141390 A1 US 2008141390A1
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seq
nucleic acid
plant
nitrogen
acid construct
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Patrick S. Schnable
Sudhansu DASH
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Iowa State University Research Foundation ISURF
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Assigned to IOWA STATE UNIVERSITY RESEARCH FOUNDATION, INC. reassignment IOWA STATE UNIVERSITY RESEARCH FOUNDATION, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: SCHNABLE, PATRICK S., DASH, SUDHANSU
Publication of US20080141390A1 publication Critical patent/US20080141390A1/en
Priority to US14/462,017 priority patent/US9523099B2/en
Priority to US15/348,425 priority patent/US10364437B2/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8222—Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8237—Externally regulated expression systems
    • C12N15/8238—Externally regulated expression systems chemically inducible, e.g. tetracycline

Definitions

  • the present invention relates to plant genes involved in nitrate uptake and metabolism.
  • nitrogen is a major component of proteins and nucleic acids, as well as various secondary metabolites found in plants (Marschner, M., Mineral Nutrition of Higher Plants, 2d ed., Academic Press Ltd.: London (1995)). Therefore, nitrogen is one of the most important inorganic nutrients of plants. Inorganic nitrogen is added to many crop plants in the form of nitrogenous fertilizers (see Frink et al., “Nitrogen Fertilizer: Retrospect and Prospect,” Proc. Natl. Acad. Sci. USA 96:1175-1180 (1999)). Nitrogen is principally added to the soil in the form of ammonia (NH 4 + ) and nitrate (NO 3 ⁇ ).
  • inorganic nutrient fertilizers are one of the major expenses incurred by producers of high-yielding crop plants (see Good et al., “Can Less Yield More? Is Reducing Nutrient Input Into the Environment Compatible with Maintaining Crop Production?” Trends in Plant Science 9(12):597-605 (2004)). Further, reports have indicated that nitrogen-based fertilizers may be associated with environmental damage (see Vitousek et al., “Human Alternation of the Global Nitrogen Cycle: Causes and Consequences,” Ecol. Appl. 7:737-750 (1997)). Therefore, one important way of decreasing the amount of inorganic nitrogen that is applied to plant crops is to develop ways to improve nitrate use efficiency (“NUE”) in plants.
  • NUE nitrate use efficiency
  • the present invention is directed to overcoming these and other deficiencies in the art.
  • the present invention relates to nucleic acid molecules from corn (maize) that are modulated by nitrogen (e.g., that up-regulated by nitrogen).
  • the present invention also relates to isolated proteins or polypeptides encoded by the nucleic acid molecues.
  • the present invention further relates to promoters of the nucleic acid molecules of the present invention.
  • the present invention further relates to a nucleic acid construct having a nucleic acid molecule of the present invention (i.e., a nucleic acid molecule that is modulated, e.g., up-regulated, by nitrogen in corn).
  • the construct also includes a 5′ DNA promoter sequence and a 3′ terminator sequence.
  • the nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule.
  • the present invention also relates to an expression system, host cells, plant cells, plants, and plant seeds having a nucleic acid construct that includes a nucleic acid molecule that is modulated by nitrogen in corn.
  • Another aspect of the present invention is a method of expressing a nucleic acid molecule that is modulated by nitrogen in a plant.
  • This method involves providing a transgenic plant or plant seed transformed with a nucleic acid construct having a nucleic acid molecule that is modulated by nitrogen in corn, a 5′ DNA promoter sequence, and a 3′ terminator sequence.
  • the method involves growing the transgenic plant or a transgenic plant grown from the transgenic plant seed under conditions effective to express the nucleic acid molecule in the transgenic plant or the plant grown from the transgenic plant seed.
  • Another aspect of the present invention relates to an isolated DNA promoter from corn suitable for inducing nitrogen-regulated expression of a protein encoded by an isolated DNA molecule operably associated with the DNA promoter.
  • the present invention further relates to a nucleic acid construct including the isolated DNA promoter, as well as expression vectors, host cells, plants, and plant seeds containing the nucleic acid construct.
  • the present invention also relates to a method of directing nitrogen-regulated expression of an isolated nucleic acid in plants. This method involves transforming a plant cell with the nucleic acid construct described in this paragraph and regenerating a plant from the transformed plant cell. By this method, expression of the nucleic acid molecule, under control of the DNA promoter, occurs in the plant and is upregulated by nitrogen.
  • Nitrate use efficiency affects both grower profitability and the ecological sustainability of intensive corn production.
  • the present invention is effective in providing a means to improve the NUE by enhancing the nitrogen uptake of crop plants such as corn.
  • the nucleic acid constructs of the present invention can be used to develop corn germplasm using marker-assisted selection and/or transgenic approaches.
  • the present invention is useful in increasing the nitrate absorption and usage efficiency by crop plants and thus reduce the use of nitrate supplements.
  • the nucleic acid constructs of the present invention include nucleic acid molecules corresponding to genes of corn plants and, hence, have the most direct bearing on nitrate metabolism in corn. Therefore, such genes may be more directly relevant to corn improvement than genes from non-crop plants such as Arabidopsis.
  • the present invention relates to nucleic acid molecules (e.g., genes) from corn (maize) (e.g., from B73 seedlings) that are modulated (e.g., up-regulated) by nitrogen (e.g., in the form of nitrate, calcium nitrate, etc.).
  • genes e.g., genes from corn (maize) (e.g., from B73 seedlings) that are modulated (e.g., up-regulated) by nitrogen (e.g., in the form of nitrate, calcium nitrate, etc.).
  • nitrogen e.g., in the form of nitrate, calcium nitrate, etc.
  • the present invention provides nucleotide sequences of the full-length cDNA clones of such genes.
  • the present invention also provides the amino acid sequences of the isolated proteins or polypeptides encoded by these genes, as well as their putative promoters (upstream of transcription start site of the genes).
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:1, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:1 has an amino acid sequence of SEQ ID NO:2, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:1 has a nucleotide sequence of SEQ ID NO:3, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:4, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:4 has an amino acid sequence of SEQ ID NO:5, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:4 has a nucleotide sequence of SEQ ID NO:6, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length EDNA clone having the nucleotide sequence of SEQ ID NO:7, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:7 has an amino acid sequence of SEQ ID NO:8, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:7 has a nucleotide sequence of SEQ ID NO:9, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a fill-length cDNA clone having the nucleotide sequence of SEQ ID NO:10, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:10 has an amino acid sequence of SEQ ID NO:11, as follows: MDRNLSGFLIGCLGAAVTLLAYQQTVVTSTQSVAAGFVVILFALFVKEGFISL.
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:10 has a nucleotide sequence of SEQ ID NO:12, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a fill-length cDNA clone having the nucleotide sequence of SEQ ID NO:13, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:13 has an amino acid sequence of SEQ ID NO:14, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:13 has a nucleotide sequence of SEQ ID NO:15, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:16, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:16 has an amino acid sequence of SEQ ID NO:17, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:16 has a nucleotide sequence of SEQ ID NO:18, as follows.
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length EDNA clone having the nucleotide sequence of SEQ ID NO:19, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:19 has an amino acid sequence of SEQ ID NO:20, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:19 has a nucleotide sequence of SEQ ID NO:21, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:22, as follows
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:22 has an amino acid sequence of SEQ ID NO:23, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:24, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:24 has an amino acid sequence of SEQ ID NO:25, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:24 has a nucleotide sequence of SEQ ID NO:26, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:27, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:27 has an amino acid sequence of SEQ ID NO:28, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:27 has a nucleotide sequence of SEQ ID NO:29, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:30, as follows:
  • a predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:30 has an amino acid sequence of SEQ ID NO:31, as follows:
  • Another predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:30 has an amino acid sequence of SEQ ID NO:32, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:30 has a nucleotide sequence of SEQ ID NO:33, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:34, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:34 has an amino acid sequence of SEQ ID NO:35, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:36, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:36 has an amino acid sequence of SEQ ID NO:37, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:36 has a nucleotide sequence of SEQ ID NO:38, as follows:
  • a suitable nucleic acid molecule that is modulated (e.g., up-regulated) by nitrogen is the non-symbiotic hemoglobin gene (MEST129-C09.T3Seq) from corn having the nucleotide sequence of SEQ ID NO:39, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:39 has an amino acid sequence of SEQ ID NO:40, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:42, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:42 has an amino acid sequence of SEQ ID NO:43, as follows:
  • a suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:44, as follows:
  • the predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:44 has an amino acid sequence of SEQ ID NO:45, as follows:
  • a putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:44 has a nucleotide sequence of SEQ ID NO:46, as follows:
  • the present invention relates to a nucleic acid construct having a nucleic acid molecule that is modulated by nitrogen in corn.
  • the construct also includes a 5′ DNA promoter sequence and a 3′ terminator sequence.
  • the nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule.
  • the nucleic acid molecules of the present invention may be inserted into any of the many available expression vectors and cell systems using reagents that are well known in the art.
  • Suitable vectors include, but are not limited to, the following viral vectors such as lambda vector system gt11, gt WES.tB, Charon 4, and plasmid vectors such as pG-Cha, p35S-Cha, pBR322, pBR325, pACYC177, pACYC1084, pUC8, pUC9, pUC18, pUC19, pLG339, pR290, pKC37, pKC101, SV40, pBluescript II SK ⁇ or KS ⁇ (see “Stratagene Cloning Systems” Catalog (1993) from Stratagene, La Jolla, Calif., which is hereby incorporated by reference in its entirety), pQE, pIH821, pGEX, pET series (see Studier et
  • Recombinant molecules can be introduced into cells via transformation, particularly transduction, conjugation, mobilization, or electroporation.
  • DNA sequences are cloned into the vector using standard cloning procedures in the art, as described by Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor, N.Y.: Cold Spring Harbor Press (1989), and Ausubel et al., Current Protocols in Molecular Biology, New York, N.Y.; John Wiley & Sons (1989), which are hereby incorporated by reference in their entirety.
  • the various nucleic acid molecule sequences may normally be inserted or substituted into a bacterial plasmid.
  • Any convenient plasmid may be employed, which will be characterized by having a bacterial replication system, a marker which allows for selection in a bacterium, and generally one or more unique, conveniently located restriction sites.
  • Numerous plasmids referred to as transformation vectors, are available for plant transformation. The selection of a vector will depend on the preferred transformation technique and target species for transformation. A variety of vectors are available for stable transformation using Agrobacterium tumefaciens, a soilborne bacterium that causes crown gall.
  • Crown gall are characterized by tumors or galls that develop on the lower stem and main roots of the infected plant. These tumors are due to the transfer and incorporation of part of the bacterium plasmid DNA into the plant chromosomal DNA.
  • This transfer DNA (T-DNA) is expressed along with the normal genes of the plant cell.
  • the plasmid DNA, pTi, or Ti-DNA, for “tumor inducing plasmid,” contains the vir genes necessary for movement of the T-DNA into the plant.
  • the T-DNA carries genes that encode proteins involved in the biosynthesis of plant regulatory factors, and bacterial nutrients (opines).
  • the T-DNA is delimited by two 25 bp imperfect direct repeat sequences called the “border sequences.”
  • control elements or “regulatory sequences” are also incorporated into the vector-construct. These include non-translated regions of the vector, promoters, and 5′ and 3′ untranslated regions which interact with host cellular proteins to carry out transcription and translation. Such elements may vary in their strength and specificity. Depending on the vector system and host utilized, any number of suitable transcription and translation elements, including constitutive and inducible promoters, may be used. Tissue-specific and organ-specific promoters can also be used.
  • a constitutive promoter is a promoter that directs expression of a gene throughout the development and life of an organism.
  • Examples of some constitutive promoters that are widely used for inducing expression of transgenes include the nopaline synthase (“NOS”) gene promoter from Agrobacterium tumefaciens (U.S. Pat. No. 5,034,322 to Rogers et al., which is hereby incorporated by reference in its entirety), the cauliflower mosaic virus (“CaMY”) 35S and 19S promoters (U.S. Pat. No.
  • NOS nopaline synthase
  • CaMY cauliflower mosaic virus
  • An inducible promoter is a promoter that is capable of directly or indirectly activating transcription of one or more DNA sequences or genes in response to an inducer. In the absence of an inducer, the DNA sequences or genes will not be transcribed.
  • the inducer can be a nutrient (e.g., nitrogen, including nitrogen in the form of nitrate), a chemical agent, such as a metabolite, growth regulator, herbicide, or phenolic compound, or a physiological stress directly imposed upon the plant such as cold, heat, salt, toxins, or through the action of a pathogen or disease agent such as a virus or fungus.
  • a plant cell containing an inducible promoter may be exposed to an inducer by externally applying the inducer to the cell or plant such as by spraying, watering, heating, or by exposure to the operative pathogen.
  • An example of an appropriate inducible promoter is a glucocorticoid-inducible promoter (Schena et al., “A Steroid-Inducible Gene Expression System for Plant Cells,” Proc. Natl. Acad. Sci. 88:10421-5 (1991), which is hereby incorporated by reference in its entirety).
  • transgene-encoded protein is induced in the transformed plants when the transgenic plants are brought into contact with nanomolar concentrations of a glucocorticoid, or by contact with dexamethasone, a glucocorticoid analog (see Schena et al., “A Steroid-Inducible Gene Expression System for Plant Cells,” Proc. Natl. Acad. Sci. USA 88:10421-5 (1991); Aoyama et al., “A Glucocorticoid-Mediated Transcriptional Induction System in Transgenic Plants,” Plant J.
  • inducible promoters include promoters that function in a tissue specific manner to regulate the gene of interest within selected tissues of the plant. Examples of such tissue specific or developmentally regulated promoters include seed, flower, fruit, or root specific promoters as are well known in the field (U.S. Pat. No. 5,750,385 to Shewmaker et al., which is hereby incorporated by reference in its entirety).
  • tissue- and organ-specific promoters have been developed for use in genetic engineering of plants (Potenza et al., “Targeting Transgene Expression in Research, Agricultural, and Environmental Applications: Promoters used in Plant Transformation,” In Vitro Cell. Dev. Biol. Plant 40:1-22 (2004), which is hereby incorporated by reference in its entirety).
  • promoters include those that are floral-specific (Annadana et al., “Cloning of the Chrysanthemum UEP1 Promoter and Comparative Expression in Florets and Leaves of Dendranthema grandiflora,” Transgenic Res.
  • a suitable promoter can also be one that is gene-specific, in that it regulates transcription of a nucleic acid molecule of the present invention.
  • a suitable gene-specific promoter gene-specific promoter (derived from MAGI93503) has a nucleotide sequence of SEQ ID NO:41 as follows:
  • Suitable promoters include those having a nucleotide sequence of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and/or SEQ ID NO:46.
  • the nucleic acid construct of the present invention also includes an operable 3′ regulatory region, selected from among those which are capable of providing correct transcription termination and polyadenylation of mRNA for expression in the host cell of choice, operably linked to a modified trait nucleic acid molecule of the present invention.
  • operable 3′ regulatory region selected from among those which are capable of providing correct transcription termination and polyadenylation of mRNA for expression in the host cell of choice, operably linked to a modified trait nucleic acid molecule of the present invention.
  • a number of 3′ regulatory regions are known to be operable in plants. Exemplary 3′ regulatory regions include, without limitation, the nopaline synthase (NOS) 3′ regulatory region (Fraley et al., “Expression of Bacterial Genes in Plant Cells,” Proc. Nat'l Acad. Sci.
  • nucleic acid construct of the present invention Once the nucleic acid construct of the present invention has been prepared, it is ready to be incorporated into a host cell. Accordingly, another aspect of the present invention relates to a recombinant host cell containing one or more of the nucleic acid constructs of the present invention. Basically, this method is carried out by transforming a host cell with a nucleic acid construct of the present invention under conditions effective to achieve transcription of the nucleic acid molecule in the host cell. This is achieved with standard cloning procedures known in the art, such as described by Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Springs Laboratory, Cold Springs Harbor, N.Y. (1989), which is hereby incorporated by reference in its entirety.
  • Suitable hosts include, but are not limited to, bacterial cells, viruses, yeast cells, mammalian cells, insect cells, plant cells, and the like.
  • the host is either a bacterial cell or a plant cell.
  • Methods of transformation may result in transient or stable expression of the nucleic acid under control of the promoter.
  • a nucleic acid construct of the present invention is stably inserted into the genome of the recombinant plant cell as a result of the transformation, although transient expression can serve an important purpose, particularly when the plant under investigation is slow-growing.
  • Plant tissue suitable for transformation includes leaf tissue, root tissue, meristems, zygotic and somatic embryos, callus, protoplasts, tassels, pollen, embryos, anthers, and the like.
  • the means of transformation chosen is that most suited to the tissue to be transformed.
  • Transient expression in plant tissue can be achieved by particle bombardment (Klein et al., “High-Velocity Microprojectiles for Delivering Nucleic Acids Into Living Cells,” Nature 327:70-73 (1987), which is hereby incorporated by reference in its entirety), also known as biolistic transformation of the host cell, as disclosed in U.S. Pat. Nos.
  • particle bombardment tungsten or gold microparticles (1 to 2 ⁇ m in diameter) are coated with the DNA of interest and then bombarded at the tissue using high pressure gas. In this way, it is possible to deliver foreign DNA into the nucleus and obtain a temporal expression of the gene under the current conditions of the tissue.
  • Biologically active particles e.g., dried bacterial cells containing the vector and heterologous DNA
  • particle bombardment transformation can be used to stably introduce the nucleic acid construct into plant cells.
  • Another appropriate method of stably introducing the nucleic acid construct into plant cells is to infect a plant cell with Agrobacterium tumefaciens or Agrobacterium rhizogenes previously transformed with the nucleic acid construct.
  • the Ti (or RI) plasmid of Agrobacterium enables the highly successful transfer of a foreign nucleic acid molecule into plant cells.
  • a variation of Agrobacterium transformation uses vacuum infiltration in which whole plants are used (Senior, “Uses of Plant Gene Silencing,” Biotechnology and Genetic Engineering Reviews 15:79-119 (1998), which is hereby incorporated by reference in its entirety).
  • nucleic acid molecule may also be introduced into the plant cells by electroporation (Fromm et al., Proc. Natl Acad. Sci. USA 82:5824 (1985), which is hereby incorporated by reference in its entirety).
  • plant protoplasts are electroporated in the presence of plasmids containing the expression cassette.
  • Electroporated plant protoplasts reform the cell wall, divide, and regenerate.
  • Other methods of transformation include polyethylene-mediated plant transformation, micro-injection, physical abrasives, and laser beams (Senior, “Uses of Plant Gene Silencing,” Biotechnology and Genetic Engineering Reviews 15:79-119 (1998), which is hereby incorporated by reference in its entirety).
  • the precise method of transformation is not critical to the practice of the present invention. Any method that results in efficient transformation of the host cell of choice is appropriate for practicing the present invention. Transfromation can also be achieved using the “whisker” method, as is well known in the art.
  • Means for regeneration vary from species to species of plants, but generally a suspension of transformed protoplasts or a petri plate containing explants is first provided. Callus tissue is formed and shoots may be induced from callus and subsequently rooted. Alternatively, embryo formation can be induced in the callus tissue. These embryos germinate as natural embryos to form plants.
  • the culture media will generally contain various amino acids and hormones, such as auxin and cytokinins. Efficient regeneration will depend on the medium, on the genotype, and on the history of the culture. If these three variables are controlled, then regeneration is usually reproducible and repeatable.
  • transformed cells are first identified using a selection marker simultaneously introduced into the host cells along with the nucleic acid construct of the present invention.
  • selection markers include, without limitation, markers encoding for antibiotic resistance, such as the neomycin phosphotransferae II (“nptII”) gene which confers kanamycin resistance (Fraley et al., Proc. Natl. Acad. Sci. USA 80:4803-4807 (1983), which is hereby incorporated by reference in its entirety), and the genes which confer resistance to gentamycin, G418, hygromycin, streptomycin, spectinomycin, tetracycline, chloramphenicol, and the like.
  • nptII neomycin phosphotransferae II
  • Cells or tissues are grown on a selection medium containing the appropriate antibiotic, whereby generally only those transformants expressing the antibiotic resistance marker continue to grow.
  • Other types of markers are also suitable for inclusion in the expression cassette of the present invention.
  • a gene encoding for herbicide tolerance such as tolerance to sulfonylurea is useful, or the dhfr gene, which confers resistance to methotrexate (Bourouis et al., EMBO J. 2:1099-1104 (1983), which is hereby incorporated by reference in its entirety).
  • reporter genes which encode for enzymes providing for production of an identifiable compound are suitable.
  • the most widely used reporter gene for gene fusion experiments has been uidA, a gene from Escherichia coli that encodes the ⁇ -glucuronidase protein, also known as GUS (Jefferson et al., “GUS Fusions: ⁇ Glucuronidase as a Sensitive and Versatile Gene Fusion Marker in Higher Plants,” EMBO J 6:3901-3907 (1987), which is hereby incorporated by reference in its entirety).
  • enzymes providing for production of a compound identifiable by luminescence such as luciferase, are useful.
  • the selection marker employed will depend on the target species; for certain target species, different antibiotics, herbicide, or biosynthesis selection markers are preferred.
  • Plant cells and tissues selected by means of an inhibitory agent or other selection marker are then tested for the acquisition of the transgene (Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, N.Y.: Cold Spring Harbor Press (1989), which is hereby incorporated by reference in its entirety).
  • the transgene can be transferred to other plants by sexual crossing. Any of a number of standard breeding techniques 10 can be used, depending upon the species to be crossed. Once transgenic plants of this type are produced, the plants themselves can be cultivated in accordance with conventional procedure so that the nucleic acid construct is present in the resulting plants. Alternatively, transgenic seeds are recovered from the transgenic plants. These seeds can then be planted in the soil and cultivated using conventional procedures to produce transgenic plants. The component parts and fruit of such plants are encompassed by the present invention.
  • Suitable plants can include dicots and monocots. More particular, suitable plants can include the following: rice, corn, soybean, canola, potato, wheat, mung bean, alfalfa, barley, rye, cotton, sunflower, peanut, sweet potato, bean, pea, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, tobacco, tomato, sorghum, sugarcane, banana, Arabidopsis thaliana, Saintpaulia, petunia, pelargonium, poinsettia, chrysanthemum, carnation, crocus, marigold, daffodil, pine, Medicago truncatula, Sanderson
  • Another aspect of the present invention is a method of expressing a nucleic acid molecule that is modulated by nitrogen in a plant.
  • This method involves providing a transgenic plant or plant seed transformed with a nucleic acid construct having a nucleic acid molecule that is modulated by nitrogen in corn, a 5′ DNA promoter sequence, and a 3′ terminator sequence.
  • the nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule.
  • the method also involves growing the transgenic plant or a transgenic plant grown from the transgenic plant seed under conditions effective to express the nucleic acid molecule in the transgenic plant or the plant grown from the transgenic plant seed.
  • the transgenic plant or plant seed is provided by transforming a non-transgenic plant or a non-transgenic plant seed with the nucleic acid construct of the present invention to yield said transgenic plant or plant seed.
  • the growing step is effective in reducing nitrogen uptake of the transgenic plant or the plant grown from the transgenic plant seed.
  • the growing step is effective in increasing nitrogen uptake of the transgenic plant or the plant grown from the transgenic plant seed.
  • the growing step is effective in increasing efficiency of nitrogen utilization of the transgenic plant or the plant grown from the transgenic plant seed. Transformation of the transgenic plant or plant seed can be achieved using Agrobacterium -mediated transformation, the whisker method, vacuum infiltration, biolistic transformation, electroporation, micro-injection, polyethylene-mediated transformation, or laser-beam transformation.
  • the present invention also relates to an isolated DNA promoter from corn suitable for inducing nitrogen-regulated expression of a protein encoded by an isolated DNA molecule operably associated with the DNA promoter.
  • a suitable DNA promoter for use in this method can be any one of the promoters described herein, including, for example, the promoters having a nucleotide sequence of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and/or SEQ ID NO:46.
  • the isolated DNA promoter can be used to prepare nucleic acid constructs as previously described.
  • the isolated DNA promoter can be operably linked to an isolated nucleic acid that either has a nucleotide sequence (or encoding portion thereof) of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and/or SEQ ID NO:44, or encodes a polypeptide having an amino acid sequence of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:2, SEQ
  • genes from corn that can be regulated by the DNA promoter of the present invention include, for example, nitrate reductase, nitrite reductase, Uroporphyrin-III methyl transferase.
  • Expression vectors can be prepared by inserting the nucleic acid construct in an appropriate vector (as described in more detail supra), and transgenic host cells and plants (including their component parts such as fruits and seeds) can be produced by transforming them with the nucleic acid construct containing the DNA promoter.
  • the present invention also relates to a method of directing nitrogen-regulated expression of an isolated nucleic acid in plants.
  • This methods involves transforming a plant cell with the nucleic acid construct that includes an isolated DNA promoter suitable for inducing nitrogenregulated expression of a protein encoded by an isolated DNA molecule operably associated with the DNA promoter.
  • This method also involves regenerating a plant from the transformed plant cell. By this method, expression of the nucleic acid molecule, under control of the DNA promoter, occurs in the plant and is upregulated by nitrogen.
  • a suitable DNA promoter for use in this method can be any one of the promoters described herein, including, for example, the promoters having a nucleotide sequence of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and/or SEQ ID NO:46.

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Abstract

The present invention relates nucleic acid molecules that are modulated (e.g., upregulated) by nitrogen in corn, to proteins or polypeptides encoded by these nucleic acid molecules, and promoters of these nucleic acid molecules. The present invention relates to a nucleic acid construct having a nucleic acid molecule that is modulated by nitrogen in corn, as well as to expression systems, host cells, plants, and plant seeds having the nucleic acid construct. The present invention also relates to a method of expressing the nucleic acid molecule that is modulated by nitrogen in a plant by growing a transgenic plant or a plant grown from a transgenic seed transformed with the construct. The present invention further relates to an isolated DNA promoter that can be used to direct nitrogen-regulated expression of an isolated nucleic acid in plants.

Description

  • This application claims the benefit of U.S. Provisional Patent Application Ser. No. 60/869,290, filed Dec. 8, 2006.
    • FIELD OF THE INVENTION
  • The present invention relates to plant genes involved in nitrate uptake and metabolism.
    • BACKGROUND OF THE INVENTION
  • Nitrogen plays an important role in various plant functions, including metabolism, resource allocation, growth, and development (Crawford, N. M., “Nitrate: Nutrient and Signal for Plant Growth,” Plant Cell 7:859-868 (1995); Marschner, M., Mineral Nutrition of Higher Plants, 2d ed., Academic Press Ltd.: London (1995); and Stiit et al., “The Molecular Physiological Basis for the Interaction Between Elevated Carbon Dioxide and Nutrients,” Plant Cell Environ. 22:583-622 (1999)). Further, nitrogen is a major component of proteins and nucleic acids, as well as various secondary metabolites found in plants (Marschner, M., Mineral Nutrition of Higher Plants, 2d ed., Academic Press Ltd.: London (1995)). Therefore, nitrogen is one of the most important inorganic nutrients of plants. Inorganic nitrogen is added to many crop plants in the form of nitrogenous fertilizers (see Frink et al., “Nitrogen Fertilizer: Retrospect and Prospect,” Proc. Natl. Acad. Sci. USA 96:1175-1180 (1999)). Nitrogen is principally added to the soil in the form of ammonia (NH4 +) and nitrate (NO3 −). However, estimates of nitrogen uptake efficency have shown that between 50 and 70 percent of the applied nitrogen is lost from the plant-soil system (Peoples et al., “Minimizing Gaseous Losses of Nitrogen,” In Nitrogen Fertilizer in the Environment, Bacon, P. E., ed., Marcel Dekker, pp. 565-606 (1995)).
  • The application of inorganic nutrient fertilizers is one of the major expenses incurred by producers of high-yielding crop plants (see Good et al., “Can Less Yield More? Is Reducing Nutrient Input Into the Environment Compatible with Maintaining Crop Production?” Trends in Plant Science 9(12):597-605 (2004)). Further, reports have indicated that nitrogen-based fertilizers may be associated with environmental damage (see Vitousek et al., “Human Alternation of the Global Nitrogen Cycle: Causes and Consequences,” Ecol. Appl. 7:737-750 (1997)). Therefore, one important way of decreasing the amount of inorganic nitrogen that is applied to plant crops is to develop ways to improve nitrate use efficiency (“NUE”) in plants.
  • Traditional plant breeding and marker-assisted selection are techniques that have been investigated for developing and identifying plants with increased NUE (see Good et al., “Can Less Yield More? Is Reducing Nutrient Input Into the Environment Compatible with Maintaining Crop Production?” Trends in Plant Science 9(12):597-605 (2004)). However, these approaches are often time-consuming and labor-intensive. An alternative approach is to use genetic engineering techniques to develop transgenic crop plants that have enhanced NUE. This approach requires the identification of genes that enhance NUE. Efforts have been reported regarding identifying genes that are regulated by nitrogen levels in Arabidopsis (Scheible et al., “Genome-Wide Reprogramming of Primary and Secondary Metabolism, Protein Synthesis, Cellular Growth Processes, and the Regulatory Infrastructure of Arabidopsis in Response to Nitrogen,” Plant Physiol. 136:2483-2499 (2004)). However, there is a need to identify genes that are involved in nitrate uptake and metabolism in economically important crop plants such as corn.
  • The present invention is directed to overcoming these and other deficiencies in the art.
    • SUMMARY OF THE INVENTION
  • The present invention relates to nucleic acid molecules from corn (maize) that are modulated by nitrogen (e.g., that up-regulated by nitrogen). The present invention also relates to isolated proteins or polypeptides encoded by the nucleic acid molecues. The present invention further relates to promoters of the nucleic acid molecules of the present invention.
  • The present invention further relates to a nucleic acid construct having a nucleic acid molecule of the present invention (i.e., a nucleic acid molecule that is modulated, e.g., up-regulated, by nitrogen in corn). The construct also includes a 5′ DNA promoter sequence and a 3′ terminator sequence. The nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule.
  • The present invention also relates to an expression system, host cells, plant cells, plants, and plant seeds having a nucleic acid construct that includes a nucleic acid molecule that is modulated by nitrogen in corn.
  • Another aspect of the present invention is a method of expressing a nucleic acid molecule that is modulated by nitrogen in a plant. This method involves providing a transgenic plant or plant seed transformed with a nucleic acid construct having a nucleic acid molecule that is modulated by nitrogen in corn, a 5′ DNA promoter sequence, and a 3′ terminator sequence. The method involves growing the transgenic plant or a transgenic plant grown from the transgenic plant seed under conditions effective to express the nucleic acid molecule in the transgenic plant or the plant grown from the transgenic plant seed.
  • Another aspect of the present invention relates to an isolated DNA promoter from corn suitable for inducing nitrogen-regulated expression of a protein encoded by an isolated DNA molecule operably associated with the DNA promoter. The present invention further relates to a nucleic acid construct including the isolated DNA promoter, as well as expression vectors, host cells, plants, and plant seeds containing the nucleic acid construct. The present invention also relates to a method of directing nitrogen-regulated expression of an isolated nucleic acid in plants. This method involves transforming a plant cell with the nucleic acid construct described in this paragraph and regenerating a plant from the transformed plant cell. By this method, expression of the nucleic acid molecule, under control of the DNA promoter, occurs in the plant and is upregulated by nitrogen.
  • Nitrate use efficiency affects both grower profitability and the ecological sustainability of intensive corn production. The present invention is effective in providing a means to improve the NUE by enhancing the nitrogen uptake of crop plants such as corn. In particular, the nucleic acid constructs of the present invention can be used to develop corn germplasm using marker-assisted selection and/or transgenic approaches. Thus, the present invention is useful in increasing the nitrate absorption and usage efficiency by crop plants and thus reduce the use of nitrate supplements. The nucleic acid constructs of the present invention include nucleic acid molecules corresponding to genes of corn plants and, hence, have the most direct bearing on nitrate metabolism in corn. Therefore, such genes may be more directly relevant to corn improvement than genes from non-crop plants such as Arabidopsis.
    • DETAILED DESCRIPTION OF THE INVENTION
  • The present invention relates to nucleic acid molecules (e.g., genes) from corn (maize) (e.g., from B73 seedlings) that are modulated (e.g., up-regulated) by nitrogen (e.g., in the form of nitrate, calcium nitrate, etc.). These genes and their promoters are natural targets for use in corn improvement. These genes can be used to improve corn germplasm with the use of marker-assisted selection and/or transgenic approaches. The present invention provides nucleotide sequences of the full-length cDNA clones of such genes. The present invention also provides the amino acid sequences of the isolated proteins or polypeptides encoded by these genes, as well as their putative promoters (upstream of transcription start site of the genes).
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:1, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACGGAAGGAGTAGAAAATATT
    GCCTGCTCCGACGACCTTGAATATTCACTGGCCATTTAATTTCTACTTAC
    AAGCCTGAATGAGCTAGAGATCCATCTGCTTCTGTACGTGCTCGTCAGGT
    ACGCTCGTAAAAAGAAAAGAAAAAAAAAGAAGAGATCGAGATCGATCTGT
    TGACGACGCCCCCGTCGCCGAT ATG GGCGACCTCTCTGTCGGCCACAGC
    CGCCGCTGGTGCGGCCGTTTCGCGGCCGTCCTTTGCCTGTGCGCGGC
    CTTCTGCAAGCCAGATGAACTCCCGATGGATCCACTGCCGAACTTGC
    CGCCGACGAGGTCGCTGCAGTGCTTCGAGGACGAACAGGTGTACAGC
    TGCTGCGAGGGCGCGTACAGGCTAAACCCATCGGGAATCATCGCCGT
    TCCCGTCGGCGCGGTGGACTACTACTGCGGCGGCGCGTGCGTGGTGG
    AGACGGAGGACGTGCTCAACTGCGTGGCCAGCGCCCTGGACGGCTTC
    GCCTTCTACAACGGGGCCTCCGTGGAGGACGTGCGCTACGCACTCAG
    GCGGGGCTGCAGCCACACCGCCAGAAGAGGCGACTTCAACGATTTGG
    AGCCGCATCTGGGCGACTACCCTGACATCTATGGCGACGATGATGAG
    CACAGCTTTGGCAGCAAGGTTGTTGCAGCTCCTCTGAGGTTGCTCGC
    GTTTCTTGGCGGTGCGGGGCTGTTCTTCCTGGGCCCTTGA

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW13E07-Full_Length cloned into pCR4-TOPO) (derived from MEST13-E07, GB_ACC# BG840928).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:1 has an amino acid sequence of SEQ ID NO:2, as follows:
  • MGDLSVGHSRRWCGRFAAVLCLCAAFCKPDELPMDPLPNLPPTRSLQCFE
    DEQVYSCCEGAYRLNPSGIIAVPVGAVDYYCGGACVVETEDVLNCVASAL
    DGFAFYNGASVEDVRYALRRGCSHTARRGDFNDLEPHLGDYPDIYGDDDE
    HSFGSKVVAAPLRLLAFLGGAGLFFLGP
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:1 has a nucleotide sequence of SEQ ID NO:3, as follows:
  • ccgcaagagggagtctttaccgagtgtcacctaatacgcttggcgaaggacctggtaaaagggcccac
    agggagctttttgctaagtgtctgtacagtggacactcggcaaagagtgagcctttgccgagtgtcac
    tccgtcaccgttacctggtgtcgtgacgacggcttttctttgtcgagtaccgagtgacactcgacaaa
    acctttgccgagcgtccgataaaaagtattcggcaaagaagccgttgccgtctttgccgagtgttttc
    cagactttgccgagtgtttcagacactcggcaaagaacctgattccgatagtgaaggtcttacacccc
    gatccaccccaattcgtgcgtattggagcaagtacccaaacaaaaccgtactgggaataattacctcc
    gttcgctgcagtttgcagaacagcagttcaatgctacaggacgacgcagctgcagcgaacatgcatgc
    atttgaactcactccgttcactgatggacaagaggcatctgggtgactaataaaagaacgacacacac
    ggacagcttctagaagtattggtagcgcatgaacaacaatgccgctgttagcttgtactgaggcacga
    aacatgaatctgacctactactgacttctactataataatagtatatagtatggccaggccaggccaa
    ctccggcgaaaacgggagtacgcatgcagatggagcggcacattagtaggctgtttggtttgaagaat
    gggctagtctatcatcttctcactctccacttttttgtttggtttgtggaatgaaatgagttgattca
    tcatcacctcattccttatagttagttagttagtactaatatgaggaatatggtcatcccaccaaatt
    tgaggaatggatccacgatgtaccaccacattttgcatgaagtgattcctcaaaccaaacacccccaa
    atgtaaaccgagtcatgcctccgatcccaaccttcgtgtttcccaccaaacacacgcgtacagaggcc
    aagcacacgcacaaaagcaagcctcgatcgtagcccgtgcctaaccctgccgatgccgtaataaactt
    gtgtgctccacgcaaccatgaaatgaacctagaaatcgcaggggcgggatgcgagtgaaaaggagcgg
    gcaggtcaggtaggtttgaactctctcctataataatcctagctagcacacttgcccagattatattg
    cctgctccgacgaccttgaatattcactggccatttaatttctacttacaagcctgaatgagctagag
    atccatctgcttctgtacgtgctcgtcaggtacgctcgtaaaaagaaaagaaaaaaaaagaagagatc
    gagatcgatctgttgacgacgcccccgtcgccgatatgggcgacctctctgtcggccacagccgccgc
    tggtgcggccgtttcgcggccgtcctttgcctgtgcgcggccttctgcaagccaggtgcgtgctcacc
    gtcaacacacgcaccattattccaccctcccaaggagcacagtacaacgcacgtacatatacctctcc
    tcaatcgatatatagttacgtcttacgtactatctagttaatctatcacgttgatgtctaatatagac
    tccgcatggcatatgcatgcagatgaactcccgatggatccactgccgaacttgccgccgacgaggtc
    gctgcagtgcttcgaggacgaacaggtaagctaacaagcaagagcgtgtttggtttcatgctaggaca
    gagttgcataccacgtagctatcataagcctaccacacgtagctatcacagcctgtcgatttcgttcg
    gtcgcctgacggtaaacatcgctgcccgagaggcgagctctttttgacaagcctcgacgaaccaaata
    agccaagtcctactgtacgagggcgatcgaggcgccgaggcctgtgtgatgtgatgccgtgtgtcgtg
    gtcacccaccagctgctgtgtacattggtccccgtgccgcgcgtcgtaaccgcatgcggcatgccgct
    gcatgcaggtgtacagctgctgcgagggcgcgtacaggctaaacccatcgggaatcatcgccgttccc
    gtcggcgcggtggactactactgcggcggcgcgtgcgtggtggagacggaggacgtgctcaactgcgt
    ggccagcgccctggacggcttcgccttctacaacggggcctccgtggaggacgtgcgctacgcactca
    ggcggggctgcagccacaccgccagaagaggtccccaagtttctcgcctactagctcatctctctcta
    cgtaccagccaagctagatcgactaccagtctccgcagcagtgcattcggaacgaccgctgacaaact
    gacaggctcgtgttcctgtcagcgcaggcgacttcaacgatttggagccgcatctgggcgactaccct
    gacatctatggcgacgatgatgaacacagctttggcagcaaggttgttgcagctcctctgaggttgct
    cgcgtttattggcggtgcggggctgttcttcctgggcccttgaacgaagatataaaagaactagcgat
    gtgatccgcgtaaatatatactccgtatatagcatgacatgagtatctagtttgtcttatatggtaaa
    ccatactaaattttcttgtatggcattaaaaaaaattaagactttatttagttatttgactagttgtt
    ctctctggatcctctaatcagttcgaactctataagcttttttattccactcctatctagaggtcgca
    taatatgctaaggtgagatcttgatgtctttcgtttttttaactcgataaagttgttgtgagtctctc
    ttataaaattatttttaatgctaatattagattttagtcagagatatgcagttgaccgttttgcacta
    aaatatttttgaatttactatagtattagttgtctactaatcacagctaaaaccgtttttatttttag
    tttttttataacagaaaaaatatctctggaaacgaaaacggcaacacagtagttcaaaaatatcgaag
    acaataatttaacatgaaaaatatatatgtaatgatcggaatctaaaaaacaatcactaaatataaac
    atatagtaacatgtactctcaattgacctgaaaaaagcacataacctatagatccacaaagtaacgaa
    gattgaagcatgaaaaatagaccatcatacattaaagggttgtgcttatttagctctagaataacctc
    cttaagagcaacttcatttgcaacaacattgtctagagttaaagagaatattttcttctctgtaaacc
    atttcaataagcatgaactgggtctaagagacaaattcttaccgttgtgccgacccagaacatgatcg
    aaatttgtaattcgcttctgtatttgtgaatcatcatctacccaatgtaccatgatacacatgtacct
    tttattctgatttgatatccacatctccatggtagcactgaagtgacaattaagggttttaaaaaatt
    tatacaacacatcatttttatgcaaaagagatccattacttcttttctaacaatgacatgtgacttta
    tagaaaatataggctttaaaggtttaatgaaatccataaagtattcatgttcaagaatgttaaatggg
    tactcatgaatgataatagtagtataaaacttcctcaaactaattgactcgtcatatttatatggttg
    gacaatatatagatatacctaccttatgatctttttctgatttgagctcctgctatcttagtaaaacc
    ttatgataacgcttctaatgcaaccaaaactaagttgttcctctatggcttttagcactacctttgta
    agtcctatttttgtagctcagaaactttcatttggcccaaatttgctccagagatttgcaattcccct
    ccactacaacaacatacaaatcaaaatactgccaaacatctaaagtatacttatttgctactttatgg
    ggtgctcatcaacattagattcact

    (>MAGI4—8075 MAGI4.contigs_w_singleton.fas 4037 bp).
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:4, as follows:
  • GGACACTGACATGGACTGAAGGAGTAGAAAATACAGAACCCTGCAACTG
    CAAGCTAAGGAGAGTGTGATCACCAACAGCTAGTGCTAGTCCCCCTTCCT
    TCCATCCATCC ATG GCATGCGTCAGCACCTTCCAGAGCTGCCCCATTG
    CCAGAAGAGCAAAGATCAACACCAGGTCCAGGGGCAGCAGCAGTAGC
    GTGGCGAAGGGGTCACCACCACCAGCCTTCCAGTTCCAGTGCAGGGC
    GTCGACTTTCGCGGCGGACACCAGCCTCCGGCTCGAGCTGGACGAGA
    ACCCCGAGGCGATCATCTCGGGGGCGTGGCCCGGGAACTGCTCCCTC
    CTCAGCTACGACGACCTCCGCGCCTACCTCGAGTCGCAGGAGACGGC
    GGCCCAGGCAGACGATCAGCGCGGCGTGGCGCTCCTGAGCGAGACC
    ATGTCCACACCCGTGCTGGTGGCCACAGCAGACCAGACCCTGGAGGA
    CGTCGAGTGCCACTTCGAGGCCGTGTCGGGGCTTCCGGTCGTCGACA
    GCGGCCTCAGATGCGTCGGGGTGATCGTCAAGAACGACCGGGCAAGA
    GCCTCTCATGGGTCCAAGACGAAGATATCGGAAGTGATGACATCTCC
    AGCTATCACACTATCGTCTGACAAAACCGTGATGGATGCTGCTGTTCT
    CATGCTCAAGAAGAAGATCCACAGATTACCAGTTGTAAACCAGGACG
    AAAAAGTAATAGGTATAGTTACCCGCGCTGATGTTCTTCGCGTGTTGG
    AAGGCATGTTGAAGATTTAGGAGCGCAGATACCCATGCTCGGAAGCCAC
    AGCCTCTTGTAAATATGTAGATGTGCCCGGGCATGGTGTTTCTGAGTAGC
    AGCAAAGAGATCTACCATGTATAGGAGTTTCTCCTTGTAAATAATAGTAG
    CACGCCAGGAGACTCCATCCCAGG

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW13A08-Full_Length cloned into pCR4-TOPO) (derived from MEST13-A08, GB_ACC# BG840889).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:4 has an amino acid sequence of SEQ ID NO:5, as follows:
  • MACVSTFQSCPIARRAKINTRSRGSSSSVAKGSPPPAFQFQCRASTFAAD
    TSLRLELDENPEAIISGAWPGNCSLLSYDDLRAYLESQETAAQADDQRGV
    ALLSETMSTPVLVATADQTLEDVECHFEAVSGLPVVDSGLRCVGVIVKND
    RARASHGSKTKISEVMTSPAITLSSDKTVMDAAVLMLKKKIHRLPVVNQD
    EKVIGIVTRADVLRVLEGMLKI
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:4 has a nucleotide sequence of SEQ ID NO:6, as follows:
  • caacgtggagtaggcaagcgttggtcttggccgaaccacgggataaaccactgtgtcaactctgtgat
    tgatctcttgtggtattgtgttttgttgagactcttttctagccacttggcatttagtgtgctaacac
    ttaacaagtttttgtggctataagtttaagttttacaggatcacctattcaccccccccccctctagg
    tgctctcaagttgacaggtgtctccatggtgcctaattgagccgttcttttacgctatccattcgatt
    ggtttggtgggtatgtgtggtctggctcttgcgatgtttgcccgctggaaacgaaacaatcgtgcata
    cgtgcgcatgcaattaacggtggtgtttttggcctgtcttgcagcagcgggtgcagcattggcttggc
    ataagcacgcaaccaaacaaagactaccttttggtcaatgcatgcgaagaatctgtacgagcgggcgt
    agacaaccaatgatgcgatataaaatttaaggattgaatcatattagaatcgagctttatttctattc
    attttcgaactaattttttaagtatcctaacttattgtgaagaaacgtaaatatttagatcccgatcc
    attaccacctctactcatacgtgaaaccaaacacgcggaatatccttctggttcaaatatgcagaagt
    caatgagcaggacttctgcttgtttgttcagtctctcaggcagggttacaggaggcaatacaagatgt
    tctccaacgattccctgaatcgttcaccccctctctcagtcctatgattcactcactcacccctcccc
    ctcttctccgtatgacaggaaatccccctagagggggagagctctaagctccccctccactaattaat
    catatttactgtgaaattacctatttgtagtgtaattaatagttagcaatgtgtattacgtattataa
    atattgtaccaatatttaaaactcaaaaaactaatgtataaaaatcaaatagtgcacttaaagtatta
    agggcagagctgaatagggggattgttggagaagtgaagaaatagggggaagaatagttgagaagggg
    tatttaaatatgaatagaaagtatgaatggagggaatgtttggagagagctcaagaacaagggacact
    gagctgcctacaacacgtggccctttttgtccctcttcttttttttctttcgcatctgctggctacaa
    gaggacacgcccttctattcgccgtatagagcagtgtctgtgaagtaaaagagaactatcctccaagg
    cttattttgagtgtattactcctggattcttgaatcttagctggtatggtatggtaaggagttacgtt
    gtccaggagattccaacttacggatccacactgaaaagtttgtattacccatttgttaggccccgttt
    caatctcacgggataaactttagcttcctgctaaactttagctatgtgaattgaagtgctaaagttta
    gttttaattaccaccattagctttcctgtttagattacaaatggctaaaagtagctaaaaaaagctgc
    taaagtttatctcgcaagattggaacagggcctatatggtcactttagagaggcatggaggtttaata
    gactatgacattcgtacgtggtcacctcaacaaactttattgtttgaccgaaccatagattgaattgt
    gtgacattgttctttgctcgtattattattaatagaaagtaaccttcttgggtgcggcccatacggtc
    ctgagcgcactaaatgaggcctcattggccgcggcccattcgatcctcaacgcactggataaagccag
    cgtggcgtggctaaaccacttcgtttggcatgggcctgtcggtgcacttgcccaaaccatgagcttgt
    accaaaactcgctagtggtagtggtattagtagtgaagaacttctgcaacttcaaactcaccgattct
    ctcgcggtcagtttggaagctaaaatatcggtggaaattagagagaatttgataagctaaaatctctt
    tattatttaaaattgaataataaataaattttaactcctccaatcttctccgtttttatgtctcccaa
    actcagtgtaccagatcatattcctttcattaaaaaaaaggtgaacaaagacgccaccttatccactg
    ccacgtgacagggggccaggggaatctcggcggccagtggcggcacgccacgccggccggtcgccccc
    gtcgctgtacaagatacccatgattggagcggggcaggtgcagagcagcaacgccacggctgcatgag
    atcaagaagctgccttcacttcgcccactgcagcatgccgtgtcgccgtcagagttgggcgcatatcc
    agataaaaaaaacttgcctgcttgcactgcagatgcgttgtttttgctaacagcaagcaggcaagtca
    gcagcctaaccttctttgatatttacagagaagatgaaaaggagaactggagagcagtagtggcagtc
    acttcactggtcaagcattcctatccacctcggcccacctccacctccctgacagtcattttgttata
    taaaacccatcaagctcccctgcaaggagatacagaaccctgcaactgcaagctaaggagagtgtgat
    caccaacagctagtgctagtcccccttccttccatccatccatggcatgcgtcagcaccttccagagc
    tgccccattgccagaagagcaaagatcaacaccaggtccaggggcagcagcagtagcgtggcgaaggg
    gtcaccaccaccagccttccagttccagtgcagggcgtcgactttcgcggcggacaccagcctccggc
    tcgagctggacgagaaccccgaggcgatcatctcgggggcgtggcccgggaactgctccctcctcagc
    tacgacgacctccgcgcctacctcgagtcgcaggagacggcggcccaggcagacgatcaggtacactt
    cgatctcgcggcttcttcagttcttgttaccattgtttacatctcctccagctcttgctaacccggcc
    tggacgggtctcctcctctgtggatatatacagcgcggcgtggcgctcctgagcgagaccatgtccac
    acccgtgctggtggccacagcagaccagaccctggaggacgtcgagtgccacttcgaggccgtgtcgg
    ggcttccggtcgtcgacagcggcctcagatgcgtcggggtgatcgtcaagaacgaccgggcaagagcc
    tctcatggggtcagcacctcgctcctctccctccacctctttctttctcatggggccagggccatgca
    tgcgcatcaagctgctagtttctcatagacaggcaaataagaacgacgtacgtccgttcagtttaccg
    gtctgtttctacttgtgacagtccaagacgaagatatcggaagtgatgacatctccagctatcacact
    atcgtctgacaaaaccgtgatgggtaatcttttttgcatcgcttttcttttcttttcttttcttttct
    gttcatgtgtgatttttaacaagttgaatctaacagtgcatgcctaacgtctacagatgctgctgttc
    tcatgctcaagaagaagatccacagattaccagttgtaaaccaggacgaaaaagtaataggtacggtg
    agtgagtgtcagaatgctcacaagccagcagagattaaaaaaaaaaactgcatgccatacacttaatt
    agtattatccttaattatcattgacaacacagagattatatgttgcaagggctaatggggttctaaac
    actgtcaacaggtatagttacccgcgctgatgttcttcgcgtgttggaaggcatgttgaagatttagg
    agcgcagatacccatgctcggaagccacagcctcttgtaaatatgtagatgtgcccgggcatggtgtt
    tctgagtagcagcaaagagatctaccatgtataggagttctcc

    (>MAGI4—31359 MAGI4.contigs_w_singleton.fas 3987 bp).
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length EDNA clone having the nucleotide sequence of SEQ ID NO:7, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAACCCTT
    CTCGCTCGGTTGCTCGGGAGCTTTCCCCTTCCTGTTCCTGAAGCTTCCGA
    CATCCGACCGCCTCCTCCTCCTCGTTCTACTCGCCGCCCCTTCTAGAATC
    ATCCAGAGGCGTGCCGGTGAAGCGCGAGAGCGGTGAGGC ATG GCGATGC
    AGACGGGGGTCGCGACCTCCAAGGTCCTCATCCTCGTCGGTGCAGGGAT
    GACGGGCTCGATCCTGCTGCGGAATGGCCGCTTATCTGATGTGTTGG
    GAGAACTCCAGGAGATTATGAAGGGTGTAAATCAAGGAACTTCTTCG
    GGTCCCTATGACATTGCACTTATTCAAGCTCAGATTCGGAATTTAGCG
    CAAGAAGTCAGAGATTTGACATTGTCAAAGCCCATTACCATACTGAAT
    GGCAAATCTGACTCGGGAGGCAGTTTATCATCCTACATACTGCCAGC
    AGCAGCAGTTGGAGCAATGGGTTATTGCTACATGTGGTGGAAGGGGT
    TGTCTCTCTCAGATGTCATGTTTGTCACAAAACACAACATGGCAAATG
    CTGTTCAGAGCATGTCAAAGCAGTTGGAGCAAGTTTCATCAGCACTA
    GCTGCAACAAAAAGACATCTAACTCAACGGCTTGAGAATTTGGATGG
    CAAAATGGATGAACAAGTAGAGGTCTCCAAAGCTATTAGAAATGAGG
    TCAATGATGTTAAAGATGACCTGTCTCAAATTGGATTTGATGTCGAAT
    CAATTCAGAAAATGGTTGCTGGATTGGAGGGAAAGATCGAGTTACTT
    GAGAACAAACAGGACGTGGCTAATACTGGTATCTGGTATCTCTGCCA
    AGTAGCAGGCGGTTTAAAAGATGGAATAAACACCAGGTTTTTCCAGG
    AAACCAGTGAGAAGCTGAAGCTCTCACATTCAGCTCAACCTGAAAAC
    AAGCCAGTGAAGGGGCTTGAATTTTTTTCGGAAAGCACCATGGAACA
    GAAAGTAGCTGACTCCAAACCAATTGCGGTGACAGTCGACGCTGAGA
    AGCCTGAGAAAACCGCTGCTGTAATGGGCACCACAGTGCACAGGTCT
    ATCAGGTTCTCATATCGGAAGGCAGGCCTTGCTTTGTGATCAAATCCT
    CTCCGCTTGAGATGCACGTGGCCTTCCTGGTTG

    (Underlinded=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW15E10-Full_Length cloned into pCR4-TOPO) (derived from MEST15-E10, GB_ACC# BG841093).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:7 has an amino acid sequence of SEQ ID NO:8, as follows:
  • MAMQTGVATSKVLILVGAGMTGSILLRNGRLSDVLGELQEIMKGVNQGTS
    SGPYDIALIQAQIRNLAQEVRDLTLSKPITILNGKSDSGGSLSSYILPAA
    AVGAMGYCYMWWKGLSLSDVMFVTKHNMANAVQSMSKQLEQVSSALAATK
    RHLTQRLENLDGKMDEQVEVSKAIRNEVNDVKDDLSQIGFDVESIQKMVA
    GLEGKIELLENKQDVANTGIWYLCQVAGGLKDGINTRFFQETSEKLKLSH
    SAQPENKPVKGLEFFSESTMEQKVADSKPIAVTVDAEKPEKTAAVMGTTV
    HRSIRFSYRKAGLAL
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:7 has a nucleotide sequence of SEQ ID NO:9, as follows:
  • taactctacaagctaagaatcaacatgtatgcaattccataataatcgggcatcatctatcactcatt
    gctaacttcagcactgaacatgatttcaagagtttttagcagaactactatgcgggtgatctcctttc
    agatgtagatggtttagaagtgtacataagcttgcaggggcttaaggaactgtttatttaatcttctg
    tgagcacgaacatccatagaagaactatctgaactgaagctaaagatttgcatgaaatggtaatttgt
    acacattaagtgcatcatgcaaacagaacgagtacacagtgaaacgatacagacctcccgagtcagat
    ttgccattcagtatggtaatgggctttgacaatgtcaaatctctgacttcttgcgctaaattccgaat
    ctaatatcaagcacgagcatgcaaagttaagtagaaatgaataattttaccgagatggaaagaagcaa
    gagaaacttctaagcagatgctgacactgagatagtgagatgtaagatgtattccatatgaggaagag
    catacctgagcttgaataagtgcaatgtcatagggacccgaagaagttccttgatttacacccttcat
    aatctcctagaaacacaaaaggtacatcattgccttaaataaacatttactaggaagtttcagagcat
    accatcaaaatctgtatgatatgtatcaggaatcactaactagtgaagcataagttatggtacgcaaa
    acttccgagtgccaattgggcgttgatgtaattttatcacatggtgttaatcacatccacatatagac
    agaatcaacgcttctagtaccccatcgccaagtcattcaaaaaatatcaggtatcagctatctgacaa
    cgctcaactatccaaaccgtatgaaagtgcgtgtaatcaaaatgaacatatttttttcggggttgggt
    gtggggggtataccgacctggagttctcccaacacatcagataagcggccattccgcagcaggatcga
    gcccgtcatccctgcgaggacaccatttcaccacgtaaggtgtcgaaacaacagccgattggggaaaa
    tagcatcaaatccgagagagatttgatgggggcgagaggtcgatggcggtgatgagaagaggacctgc
    accgacgaggatgaggaccttggaggtcgcgacccccgtctgcatcgccatgcctcaccgctctcgcg
    cttcaccggcacgcctctggatgattctagaaggggcggcgagtagaacgaggaggaggaggcggtcg
    gatgtcggaagcttcaggaacaggaaggggaaagctcccgagcaaccgagcgagaagggtgcctggac
    ccgggaccgggacctgagaatttcgtgtgtcacaaacaaacagggtgaaccagttgtgaaatgggacc
    acgtgtcagtgaagaggtgagtagtagtatttgtgagttgtgactcgagaaatgccgctgcgggctgc
    ggcctagccacagccacgtcagcaatgtcgaaagtcgaaaccaaccccactccacgtctcccccagga
    gaagcgaccattcaaagccgccgggagctcggcgtcaccgccgcgagctcgacacctcgacacctcgt
    gccgccgcagcgcttgctttcgtcccccttacgccactcccacttggccacttcagccaccatctccc
    tgaagctagtggctaacctcctcaccgccatgggcacccctctcctcatcccccttctcgtcaccctc
    cagctgttcactacctcctcccccgcggtcgcgtcgtcacacatctccgccatcatctcgcagtcggg
    cctcgacttcgccaaggacctgctcgtatcccatgccgttgcgaccctcacgcccatgaacgtgccgg
    acatcgagaggaccatgagcatacccctcgtgggcaccgtccgcatggccgcatccgggattgtgctc
    cacggcctcgccgtcaccaactccaccgtcgctgtgggggacgcgggtgttgtcgtggccgcctcgtt
    ggccagcgcgaacctcaccatggagtggaactactcgtatgacgcctggattgtgaccatatccgaca
    gcgggaatgcttcggtccaggtataaatgaggggaacatatactgtgcagtcatattagtgcaaccgt
    gcaattaagcaatgatgcatcgatccaatcaaaatccaactatgattgctattttaggtggaacatgg
    ttagatgcaaaacagtcctgtttggttgatattcgatattccatcagttatgttccccaaggcgtggc
    ttgctgattggtggctgttaattgaatcataagatactgcccgtttttttaatatactgagtaggaga
    tatacgcatcttttatgctattaagtatagactgatcgcgcgacacttgaattttggaatatctattt
    tctgtcagatgtcagaagtagaatcaattatcttagaagtgggtgctaattcacacctattactatat
    ttaaaatgggattaatataaacactctatttttctcgaaagcgcaagagagctgcgcgaaaatatatt
    aagaagaagtaaaaggtccaaaaggaccccaagatacagataaggccgacctacggcggccaataaca
    agcataaatgaaaccatccatgacaaaaacactgctaccagaacagcactacatctatctagctaaca
    ggtagacctgggataggggcagtaagcaaggacagcctctttgcaccagccataacccaaagatcaat
    ctccaaaccaacacttctaatagcaacagcaacactaggactcttattgtcaaaaacgtagccattgc
    gatgtttccaaagggtccaaacaccaagaatgacaagagaattaagaccatttcttgcaatcccagga
    gtcttggtgatcaagtcttgccaccaatccataaaaacctcttcacaggactgatgggccaagtgttg
    tagatttacaagaagaagcagcttgaaccaaaattctctagcaaaaacgcagcccagcagcatatgat
    ttaaggtttcctgatcctgatcacataatggacatctctccggatgatccatacctcttctttgcaac
    ctatcagctgtccacaccttcttgtgagcgaccaaccacatgaaaaatttagatttcggaggagccca
    agtcttccaaattatatgaaaaggctcaaactcaattgacccaataaagaaacccctataagcttcct
    tggaagaatattttccattggcagcaaggcgaaagaaatgcttgtcttcaacatgaggtcttagctga
    accaaatctaataaatcccacaagaggagatactcgttgatacacccactgaa

    (>MAGI4—20155 MAGI4.contigs_w_singleton.fas 3385 bp).
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a fill-length cDNA clone having the nucleotide sequence of SEQ ID NO:10, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAAAACTC
    CCAAATCCTTCGTTTCGTCGTCTCCACACGCAATAGCATCCGAGCAAAGA
    AGCCAAAGAGCAACTGGGAGCGAGGACGGGAGGCAACAAGCGGCGGCG
    GC ATG GACCGGAACCTGAGCGGGTTTCTGATCGGGTGCCTGGGCGCC
    GCCGTGACGCTGCTGGCGTACCAGCAGACGGTGGTGACCAGCACGCA
    GAGCGTCGCGGCGGGCTTCGTCGTCATCCTCTTCGCCCTCTTCGTCA
    AGGAAGCATTCATTTCCCTCTGAATCTCTGGTGCGCGTCAGCCAGCCAT
    GCATGAGGAGGCGTCATCGCTCCGCTGCCTGTATTTCTGCTCGCTAGTTC
    AGTCCCGCAGCTGCCGCTGTGCTCGTCAGGTTC

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW28B08-Full_Length cloned into pCR4-TOPO) (derived from MEST28-B08, GB_ACC# BG842208).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:10 has an amino acid sequence of SEQ ID NO:11, as follows: MDRNLSGFLIGCLGAAVTLLAYQQTVVTSTQSVAAGFVVILFALFVKEGFISL.
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:10 has a nucleotide sequence of SEQ ID NO:12, as follows:
  • gatacgactctcgctggtatataaaatctgtttcgtagataaacatgaaaccagaatttttgatcacc
    atatacttgtttcagaagcaaattggggacaccatatacttgttgccttcaaacgaccgtacaataag
    ttcagactgaccatctgaatgtcacaagagctagtttagagcagcaagaaattgtcaagtgacctaga
    catcccgaaccgacgcttcccagacttagcccgaccttccgggtccttcataagctgactccgtggcc
    cctcaccagaccaacgccgcagccgttgaccttgcggctttttatccccatccggccatccccaaccc
    aactcccaaatccttcgtttcgtcgtctccacacgcaatagcatccgagcaaagaagccaaagagcaa
    ctgggagcgaggacgggaggcaacaagcggcggcggcatggaccggaacctgagcgggtttctgatcg
    ggtgcctgggcgccgccgtgacgctgctggcgtaccagcagacggtggtgaccagcacgcagagcgtc
    gcggcgggcttcgtcgtcatcctcttcgccctcttcgtcaaggaaggattcatttccctctgaatctc
    tggtgcgcgtcagccagccatgcatgaggaggcgtcatcgctccgctgcctgtatttctgctcgctag
    ttcagtcccgcagctgccgctgtgctcgtcaggttcttggaaaaatactgtaatagcgtagtgacttt
    tatgtacgacacggatggttgttgctggctgaagggtctactctgtcgaaatcgatgtatcttagttt
    atgctacttgaagaacagcagactgcagatcagcagagttcttgccttcttacgctaattaataatta
    ttggtacacgaatcctgattgtgttgagccttcttgccgttgctccttccctactaacatctcggctt
    gccaattcacctatgtatgtttgctttgtatattagtgcaggtattaatggccgcctgtaagtgagtt
    tgttctcccttgttgaactaataaaattggcatgaattcaccccaaaaagattgatgctgtttctcac
    tagttttcagcctcagacgactatagatgtccaaacagtgcggaccgtccatttgaaacttgacccgt
    cacgattttagtccggtccaagcatggccaagcagggttggtaacggcacgacctgtttagcgtgccg
    ggtttgggcagctacagaggcccgcgtgttttggtccgatccgacacgagcaatgggccgacacagcg
    gcggcccatttttcatatggcatatggtgccagcggccacacgcccccccaaccaggccacacacccg
    aaccctatctctaatcccctcaccccctcgggccctccgtccccatctctagcgattcggcgccgtcg
    ttctcgcccgttgcatcccgtcggctcttgacctcgacggcggacgactctccatcgctgtcgtatgt
    ggtgctccgacctgcttggacttggagttcctccgtcctccctcgtcactccctccgtctgcgactgg
    ggactccctaaccctaacccctccggtctccggattcggtggttctagctcctcagctgtgcaaggtt
    cgtttatctcgtctaatcccctccagatttggtgtctagctgatgtctggtgctcgtctgtggtgtct
    ggttgccgttgccggtggtcgtcacctgttgctcct

    (>MAGI4—8905 MAGI4.contigs_w_singleton.fas 1736 bp).
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a fill-length cDNA clone having the nucleotide sequence of SEQ ID NO:13, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAATCTTG
    GATCTGGTGGTGGGTTCATCCTTGGCCCACTTCTTCTTGAGCTTGGGTGC
    ATACCGCAGGTGGCAAGTGCAACAGCAACATTCGTG ATG ATGTTCTCCT
    CCTCCCTCTCTGTGGTGGAGTTTTACTTCCTGCACAGATTCCCCCTGCC
    TTTTGCTGGCTACCTCATCTTCATTTCCATATTGGCTGGATTCTGGGG
    CCAGTGTTTGGTTAGGAAGATCGTGCATGTGCTCAAGAGAGCATCGC
    TTATTGTCTTCATCCTCTCCTCTGTTATCTTCGTCAGTGCTCTTACGAT
    GGGTGTCGTTGGAACCCAGAAGAGCATTTCGATGATCAACAATCACG
    AATATATGGGGTTCCTCAACTTCTGCGAGTAACTCAAACACCATCAGAC
    TGTCGATCCGTCCGGGAGAATCCAGGCCAATGCCTAATTGACCTCATCTC
    CCTCAAAATCTAGAAGAATAAAGTCGCCGAGTATGTGCACAAGTTAGCTC
    CTCGCCAACATGTGCGCATTTAGACCGACAGAGTCGCTGTAGTGAATTCA
    GCTCGTGTTAGCTCCTGGCTAACGAGCTGACCATACGGCTTTAGTTTTGT
    GAAGTGGGCGCGATTTCGTCATGTCATGCATGTGTTAGCTCCTGGCTAAC
    CTGCAAATGCGTGTGTTGGTGCAGGTTTTTGTCACGTCTGCGTCAGCTCC
    TGGCTGACCAGCAGTTGTTTGTCGTTCATTCTCTGCGTCAGCTCCTGGCT
    GACC

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW31A10-Full_Length cloned into pCR4-TOPO) (derived from MEST31-A10, GB_ACC# BG842452).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:13 has an amino acid sequence of SEQ ID NO:14, as follows:
  • MMFSSSLSVVEFYFLHRFPLPFAGYLIFISILAGFWGQCLVRKIVHVLKR
    ASLIVFILSSVIFVSALTMGVVGTQKSISMINNHEYMGFLNFCE
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:13 has a nucleotide sequence of SEQ ID NO:15, as follows:
  • cttgggcggtagagctttttattagcttttcaaaaagttcaaggtcatcaaggtcagagtttaaatct
    aaaagctatgcctaaaatataaaatgggtcatactgagcacccatacatatgatgatcttgtccagta
    ccacatctgatacacacagagcattacggtgacccatgttccatatcttaaggtaacgaaggtttgtc
    ctaagttaaagttttgaaactttgacaacaatatctacaaaaataattattttttacttaaagaaatt
    atatattgtgctagatgttttaataataaatataatagttttatttttatttagtcaatgtttatgaa
    tattttgttattaatgataaaagtttcaaaattttgacttagtataaactttcgtgatcttaagatag
    ggaagagagggagtgagtaggtatcaattgcacccaggtaatgatcattttcaacggtcaaattacta
    aaaatagccgttaccaaaaactcaacagtgtacatgatgtggagcgatccggggggagacacccactt
    acgttcaatgaaaatgctagtccacgaaggagacggaagcccaccctggcctctctttgaggcgaagc
    cacgttccggccaatcgtctcacagcctctatgcaggctggaatgtcacccatgctgcacctcacctc
    aaccatcgtaaatcttaaggaccattcttcttaattaactcatttgcaagggtttgtagcgccgcttt
    accttagtacatgtgttacagtaaacaaacaattgccagtgctttatatgatttcgatccatcatatt
    ttaggtccaaaacagcatcttcactcaaagagacagattaaagctgtttggactgctttagctataat
    aaaaatactgtagaaaaaacagaagtcggtggaagccgcagcgaacatgttctgattttcacggaaat
    acggcttgaaacgcactcggcttgcacaaacagaatgggaattgactgatatttacaatgttccatgc
    aacaaatatttgcagttttgcagcctagcctggtgctagcgcaagaatgaacaacaaataactgctgg
    tcagccaggagctgacgcagagaatgaacgacaaacaactgctggtcagccaggagctgacgcagacg
    tgacaaaaacctgcaccaacacacgcatttgcaggttagccaggagctaacacatgcatgacatgacg
    aaatcgcgcccacttcacaaaactaaagccgtatggtcagctcgttagccaggagctaacacgagctg
    aattcactacagcgactctgtcggtctaaatgcgcacatgttggcgaggagctaacttgtgcacatac
    tcggcgactttattcttctagattttgagggagatgaggtcaattaggcattggcctggattctcccg
    gacggatcgacagtctgatggtgtttgagttactcgcagaagttgaggaaccccatatattcgtgatt
    gttgatcatcgaaatgctcttctgggttccaacgacacctgaaactcaccgaaacaagaggccattag
    gagagaagttaaaaatcaaactagattgatttagacgaaacaagtaaaagagctaatataatgctaca
    tccgttctcgaatatttgtcgtccgttagttcattttttaaaatgaactaaaacgtgacaaataaaaa
    agaacggagaatggagtgagtattccttaagattatttttctcaaggatgcatgctataattgcaaaa
    tcaatttaagcaacaccggtacgtttagttcaatttaagcaacaccggtacgtttagttcaattcaac
    ttggagcggtatcaggttagcaatttgccaagtttaaagctaagtagcaagtcaatgagttatcaata
    ggttcatacccatcgtaagagcactgacgaagataacagaggagaggatgaagacaataagcgatgct
    ctcttgagcacatgcacgatcttcctaaccaaacactggccccagaatccagccaatatggaaatgaa
    gatgaggtagccagctgcagatagagaaacagtgcaagttattaactcgttaccatataacaatcaca
    cttatgaaaacgtctacattttgaggaattggaatctaactaatagagtaggttatttctttagaacg
    tgacatttcataa

    (>MAGI4—154269 MAGI4.contigs_w_singleton.fas 2189 bp).
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:16, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAAAAAGT
    GCTCCCGGAAGACTCCAAGCTGCAGCTACCGGCCTTCCTCTCCCCCATTC
    CAATTCCGAGAACAGGGGCGGCGGAGTCAACCAGGTACG ATG TGCTCGG
    TAGCGAGGCTGGCGTTTGTGCTTGCACTGGCCATAGCCGCCTCGTCAA
    TTGAGGTTGCGGAGAGCAGAGATTTTAATATCTTTGCTCAGGGCAGC
    TTGCCTGATGCAACCAAGGGATCGTCTGGTCTAGCTGCAACCAGTGG
    AAAGTTGTGTCAGTTATGCGAGCAGTACTCATCCGAGGCGCTCCTCTA
    TCTCACACAAAACGAGACCCAGACTGAGATTCTTAGCATTCTACACCA
    TGAATGTGCCAGCCTTGCCCCTCTCAAACAGCAGTGCATCACGCTGG
    TTGACTACTACGTACCCCTTTTCTTCTTGGAGGTCTCCATGGTTACCC
    CTGAGAAGTTCTGCGAGTCGATGCATCTCTGCAAGAAGGGGATGAAG
    ATTAGCCTACCCACCCGGGAGGGTACTTGTGGTTTGTGCCACCATGTT
    GTTGTTGAAATTCTTATCATGCTTAAAGACCCCAACATGCAGCTGGAA
    GTAATCGACCTACTCACCAAAACATGCAGCAAGGCGCAGAACTATGA
    ACAGTAGTGCAAGCGGCTGGTCCTCAAGTATATTCCACTTATTCTGGTGA
    AGGGCCAGAAATTCCTTGAGACAACGGATGTCTGCTCTGTGATACATGCA
    TGCAAAGCAGGCACACAAGCATCAATGGAAGCCATGCCTCTGTCTGCCAT
    GTTGTGAAGGTGATGCGA

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW42B12-Full_Length cloned into pCR4-TOPO) (derived from MEST42-B12, GB_ACC# BG873755).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:16 has an amino acid sequence of SEQ ID NO:17, as follows:
  • MCSVARLAFVLALAIAASSIEVAESRDFNIFAQGSLPDATKGSSGLAATS
    GKLCQLCEQYSSEALLYLTQNETQTEILSILHHECASLAPLKQQCITLVD
    YYVPLFFLEVSMVTPEKFCESMHLCKKGMKISLPTREGTCGLCHHVVVEI
    LIMLKDPNMQLEVIDLLTKTCSKAQNYEQ
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:16 has a nucleotide sequence of SEQ ID NO:18, as follows.
  • gcactcatagcacatctgaggttccctttcttgaacttagctcacctactgttcatagttctgcgcct
    tgctgcatgttttggtgagtaggtcgattacttccagctgcaagcttgcagcaaacaaagaaaggcat
    tacagtatgtacagagtacagagcagtacaacacagaagaatgttggtgacagatagtgaaaatatgg
    ttattacctgcatgttggggtctttaagcatgataagaatttcaacaacaacatggtggcacaaacca
    caagtaccctcccgggtgggtaggctaatcttcatccccttcttgcagagatgcatcgactcgcagaa
    cttctcaggggtaaccatggagacctccaagaagaaaaggggtacgtagtagtcaaccagcgtgatgc
    actgcagcagggtgaatcatcaacacaacatttaacacagctgaaaacgtggtaccaatggaaggatc
    acaagttacctatacctgctgtttgagaggggcaaggctggcacattcatggtgtagaatgctaagaa
    tctcagtctgggtctcgttttgtgtgagatagaggagcgcctcggatgagtactgctcgcataactga
    cacaactttccactggttgcagctagaccagacgatcccttggttgcatcaggcaagctgccctgagc
    tgaattgaagacagaagaaaggattggccagaaatgcaaaacttcagaaaaacttgagttcctgtgag
    gaatagcagctaagctgaagctacgccctctacattgagtagaactgatggcttagacgtaattgctt
    tctttaacatgtcaccggactaaatgaagatacgaacttgtcaaacaaagaaggaatttagataaact
    aattgaaactatcacgagatctccatcgaaaagaaactatcactagacctgataattcactgctatgg
    atcaacattcaacaaagaataaagagagtaaggagcaaaaatcagtagattgaaagcttaccaaagat
    attaaaatctctgctctccgcaacctcaattgacgaggcggctatggccagtgcaagcacaaacgcca
    gcctcgctaccgagcacatcgtacctgcttaccactaccagttggtcagttgacaggaacaaaactac
    tgcttgaagaaaactatcgcagtgaaatcagctgtggctgatggacgcagaaaagctggcttgctcaa
    agcttctccataaagccaaaaggtaaccaaaaaaaaaagagaaaggaaatgtatcctagggccctctc
    tctacgtcatgtaacggatcagtagaagtttcagattcattcagcccgacgtaactgaagaattcagt
    tcgcttcaagatgtagccatcagattcacgtatttggagtcaagccaagatagtaccaattggtccgc
    atccacattccaggcaacagattcacgagattcagctcgctccacgccagcagagctgctactattct
    ggcaccactccaaatacgcctttgcagcagattagcaaagcattttacgctcgcttttgcgctttatt
    ttgcccctcgtttcctttccaggtagcttccggttccgaagaatcggaggtccttggattcagggaca
    aggggtcgaactgggcagcaaatcaagaaccgaggggagacggtagtacagagagcccaggagaagct
    aacatatgaatggggaattaaagacgcatctcacctggttgactccgccgcccctgttctcggaattg
    gaatgggagagaggaaggccggtatctgcatcttggactcttccgggagcactttgttttcttaaagc
    ttcgtgttacattaagaagatgcatgagcatgtagaacagtgtgtgtggccgtgtgtgtgagaacctg
    agatatttttgcttctttggtggccaagatgtgttagaaaggcataatcttttctca

    (>MAGI4—114997 MAGI4.contigs_w_singleton.fas 1961 bp)
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length EDNA clone having the nucleotide sequence of SEQ ID NO:19, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAATATAG
    CCAGAACTCTTGCATCCTGGTGATGGTAAACTGCCGTGCCAGTATAAACG
    CGAAGGCAGGTCACACATACTCACAAGTCCGTCCCATCTCAGGTCATCCA
    TCCATCCATCCCTGCAGCA ATG GCGTCTGCAGTGACCAGCAGCGACAA
    GGAGCAGGCCGTCCCTACCATCGACGCTGACGAAGCGCACGCGCTGC
    TGAGCTCCGGCCATGGCTACGTGGATGTCAGGATGCGGGGGGACTTC
    CACAAGGCGCATGCGCCCGGTGCTCGGAACGTTCCCTACTACCTGTC
    CGTCACGCCGCAAGGGAAGGAGAAGAACCCACACTTTGTAGAGGAAG
    TGGCTGCCTTCTGTGGGAAGGATGATGTCTTCATTGTGGGTTGCAAC
    ACGGGGAACAGATCCAGGTTCGCGACGGCAGACCTTCTGAACGCGGG
    GTTCAAGAACGTGAGGAACCTGCAAGGTGGTTACCGCTCCTTTCAGC
    AGCGAGCTCAACAGCAGTAGACATCACGTCCTGAAGGTATGCCAGGGAT
    GCTGCAGTTGAACG

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW43D12-Full_Length cloned into pCR4-TOPO) (derived from MEST43-D12, GB_ACC# BG873856).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:19 has an amino acid sequence of SEQ ID NO:20, as follows:
  • MASAVTSSDKEQAVPTIDADEAHALLSSGHGYVDVRMRGDFHKAHAPGAR
    NVPYYLSVTPQGKEKNPHFVEEVAAFCGKDDVFIVGCNTGNRSRFATADL
    LNAGFKNVRNLQGGYRSFQQRAQQQ
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:19 has a nucleotide sequence of SEQ ID NO:21, as follows:
  • cgcactaagaaggcagggaattgtgtggcaaatataggtacatgctacacgtgtgatatttcccgatt
    tgtcaatctgggacatgaagttaacatcgcaaattataatgttacaggaaccaggtatggtgctagct
    tgcgtaagcaaatcaagaagatggaggtatctcagcattccaagtacttttgcgagttctgtgggaag
    gtacatttctgttagttaccctgtttctgcatccaagttttctaatctttgatctattgaactgcgag
    ctgtctttatgttgtactcgttatcatcaccactgctgttatgaaatgtaggctgcagtcagattgat
    tttgagcacatgaaatcaattagttttcgatatatctgtttgtcacaagcacatgaaatcaattagtc
    ttcgatatatctgtttgtcacatttgaatgatttataagatgtctgggcatgtccatcaatgtgtttc
    taagatacatttgaagacagacagcatttgttccgaatccaacctttgctgtgctgtgtttccagttt
    gctgtgaagaggaaagcagttggaatttgggggtgcaaggactgtgggaaggtgaaggctggtggtgc
    ttacaccatgaagtaagtaattcttcgcctgtccgaaaaccacaatttgttagccacggctaaattct
    gttaatgtgtttgcagcactgctagtgcggtcaccgtcaggagcacgatccgccgcctgagggagcag
    actgaagcatgatatagctctttatattattggggtttcctgtagttgctcttgtcaggcatgttgtg
    ggggccttatctagtggaaatgtggaatcactgtactggctgttttgccgagacaatgctccttatat
    ttggtttatgctctaggatctcaaagttgtgttaagatttgcccttggttaccgttctgaatctgaca
    agtgatatttcatcctatgccatcttgacgtcgaatttggttgtggttttctatgcgcttggctgtgt
    caatggtttgctattctgttcttgaaattctacagatactgctgcgtctctgctggttgagtctggtt
    tagatagcaaccagtccttattattggtctttcaagttcaagtcaactaaaatgcgacaaataaaaaa
    aagaatggagggagtatataactgttcaagtcaaccaatccttattacgcctgcacttgtgtccaaaa
    agaaatgccccggagctattattggtctgttgccagataagcagtgacgacgcagcatcgaaggtcag
    agacgacttttttgcgagaacgagcatcaagctgacggaatggagcattattccgataaaaaaaaggt
    atagccagaactcttgcatcctggtgatggtaaactgccgtgccagtataaacgcgaaggcaggtcac
    acatactcacaagtccgtcccatctcaggtcatccatccatccatccctgcagcaatggcgtctgcag
    tgaccagcaggtaaacatagcttctgagtgcatctgatgttgcttacagtaacattacatgcatagag
    cagaagatcggatgcatctggattaaccagagtcagtcttgtcttggtgtgcactgcagcgacaagga
    gcaggccgtccctaccatcgacgctgacgaagcgcacgcgctgctgagctccggccatggctacgtgg
    atgtcaggtgcgtagagctcagccagtcagggacgcgcctatgcgtgtgctggagcttccagacgaac
    tgacgctgacggggacgaggtggttctccttcgtgcaggatgcggggggacttccacaaggcgcatgc
    gcccggtgctcggaacgttccctactacctgtccgtcacgccgcaaggtcagtttcttgctcgctggc
    gttggcgctggcactggcattggggttattgatttgagctgcctctgtccccgtgtagggaaggagaa
    gaacccacactttgtagaggaagtggctgccttctgtgggaaggatgatgtcttcattgtggtagcta
    ttcactcatataaataaataaataaatgtactagtactctataaatagatagatacgcctgtaatcaa
    ggagttgtcgtgtagggttgcaacacggggaacagatccaggttcgcgacggcagaccttctgaacgc
    ggtaaacacagcccatccgagctttagcatcaatccagttagctgtatgtgtgtgtgtgtgtgtgtgt
    gtttaactgagggtcacactagtctgctcgcat

    (>MAGI4—143540 MAGI4.contigs_w_singleton.fas 2277 bp)
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:22, as follows
  • AAGATAGCTGCAAAACAAGCGAGTTACTTACAACCAAACAGAAGGGTAGA
    AACCACCTGAAGCC ATG TGCATTGCTGCATGGATTTGGCAGGCTCACCC
    TGTGCACCAACTCCTCCTGCTTCTCAACAGAGATGAGTTCCACAGCAG
    GCCTACAAAAGCAGTAGGATGGTGGGGTGAAGGCTCAAAGAAGATCCT
    TGGTGGCAGGGATGTGCTTGGTGGAGGAACATGGATGGGGTGCACCA
    AGGATGGAAGGCTTGCCTTCCTGACCAATGTGCTTCAACCAGATGCCA
    TGCCCGGTGCACGGACTAGGGGAGATCTGCCTCTCAAATTCCTGCAGA
    GCAACAAGAGCCCACTCGAAGTTGCAACTGAAGTGGCAGAAGAAGCTG
    ATGAATACAATGGCTTCAACCTCATACTAGCTGATCTAACAACAAATAT
    CATGGTTTATGTGTCAAACCGGCCTAAGGGTCAGCCTGCAACAATTCA
    ACTCGTGTCACCAGGACTCCATGTGCTGTCCAATGCAAGGCTAGATAG
    CCCTTGGCAGAAGGCAATTCTCCTCGGTAAAAACTTCAGGGAGCTTCT
    TAGGGAGCATGGTGCTGATGAGGTTGAAGTGAAGGATATAGTTGAGAG
    GCTAATGACTGACACCACAAAGGCTGACAAAGATAGACTGCCAAACAC
    TGGTTGTGATCCCAACTGGGAGCATGGTCTGAGCTCCATCTTCATTGA
    GGTGCAAACTGACCAAGGGCCCTATGGGACACGGAGCACAGCCGTTTT
    ATCAGTGAACTATGATGGCGAAGCTAGCTTGTACGAGAAGTATCTTGA
    GAGTGGTATATGGAAGGATCACACAGTGAGTTACCAGATAGAG TAG TA
    GGCATTGCACAGGAAAAGTTGGCGACCTCA

    (Underlined=start and stop codons; coding sequence in bold) (Sequence of 5′ RACE product AM45C08-1T3 Full_Length cloned into pCR4-TOPO).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:22 has an amino acid sequence of SEQ ID NO:23, as follows:
  • MCIAAWIWQAHPVHQLLLLLNRDEFHSRPTKAVGWWGEGSKKILGGRDVL
    GGGTWMGCTKDGRLAFLTNVLEPDAMPGARTRGDLPLKFLQSNKSPLEVA
    TEVAEEADEYNGFNLILADLTTNIMVYVSNRPKGQPATIQLVSPGLHVLS
    NARLDSPWQKAILLGKNFRELLREHGADEVEVKDIVERLMTDTTKADKDR
    LPNTGCDPNWEHGLSSIFIEVQTDQGPYGTRSTAVLSVNYDGEASLYEKY
    LESGIWKDHTVSYQIE

    (The above sequences are presented after trimming GeneRacer Oligo sequence. Cloned in pCR4-TOPO wctor at the “TOPO Cloning site”.)
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:24, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAAATCAG
    CCGCAGTCGCGTCGCGTCGCGTCGCGTCCAGTCCAATCCTCGGAGCCTCA
    CACGGGCGGACGAGCGGGAGCTTCTCCCAATCTCCCCTGCCCTGCCCTGC
    CCTGCCGCCGCGCTTAGCTTCGCATCTTCCCCTCCTCCTCCTCCTCCTTC
    CTCGGCCAAGCGAGGAGCGAGGCGCGGGCGCGAGCGCGTCGTTGAG ATG
    GATTCGGAGGCGGTGCAGCACGGCCTTCTCCCTCTGTCTGCCTGTCCTCC
    TACCGCCAACAGCTGCGCGCATTACAGCCGTGGGTGCAGCGTCGTGG
    CGCCCTGCTGCGGCCAGGCCTTCGGCTGCCGCCATTGCCACAACGAC
    GCCAAGAACTCGCTGGAGGTCGACCCGCGCGACCGGCACGAGATCCC
    CCGCCACGAAATAAAGAAGGTGATCTGTTCTCTCTGCTCCAAGGAAC
    AGGACGTGCAACAGAACTGCTCCAGCTGTGGGGCCTGCATGGGCAAG
    TACTTCTGTAAAGTATGCAAGTTCTTCGATGATGATGCCTCAAAGGGC
    CAGTACCACTGTGACGGATGTGGAATATGTAGAACCGGCGGCGTGGA
    GAACTTTTTCCACTGTGATAAATGTGGGTGTTGCTACAGCAATGTCTT
    GAAGGATTCCCACCACTGCGTCGAAAGAGCAATGCATCACAACTGCC
    CCGTCTGCTTTGAGTATCTGTTCGACTCCACGAAGGACATCAGCGTGC
    TGCAATGTGGGCATACCATCCATTTGGAGTGCATGAACGAGATGAGA
    GCACACCATCACTTCTCATGCCCAGTGTGCTCGAGGTCCGCCTGCGA
    CATGTCGGCCACATGGCGGAAGCTCGACGAGGAGGTCGCGGCCACG
    CCGATGCCTGACATCTACCAGAAGCACATGGTGTGGATCCTGTGCAA
    CGACTGCAGCGCGACCTCGAGCGTGCGGTTCCACGTGCTGGGGCACA
    AGTGCCCCGCGTGCAGCTCGTACAACACCCGGGAGACGAGGGCTGCG
    TGCCCCAGGATCTGAGGCGAACCAGAGGCCATGTCACAAAATGCCAGGG
    AGATGCCGTCCAACGACCATCTGTCTGCAGGACGTTGCTGCGCTTAAGGT
    TAAAGGCTAGCGCGAGACCAGGCCTGGTAGTCCAGTCTTGAGTTTGGTGC
    TGGAGCATTTGTAATGTTCCGGTAAAATGTAATGCGTCCATGAGTGCTGT
    CCAGGCAGTAAGCACACCTGTGGATCGGGGCCGGCGCAAGGTCCCTAGGC
    AAGCTGCAGGATTAGTGGGGCTATTCATGTTTAGGGCGCGAATGCAACGA

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW55C10-Full_Length cloned into pCR4-TOPO) (derived from MEST55-C10, GB_ACC# BM072886).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:24 has an amino acid sequence of SEQ ID NO:25, as follows:
  • MDSEAVQHGLLPLSACPPTANSCAHYSRGCSVVAPCCGQAFGCRHCHNDA
    KNSLEVDPRDRHEIPRHEIKKVICSLCSKEQDVQQNCSSCGACMGKYFCK
    VCKFFDDDASKGQYHCDGCGICRTGGVENFFHCDKCGCCYSNVLKDSHHC
    VERAMHHNCPVCFEYLFDSTKDISVLQCGHTIHLECMNEMRAHHHFSCPV
    CSRSACDMSATWRKLDEEVAATPMPDIYQKHMVWILCNDCSATSSVRFHV
    LGHKCPACSSYNTRETRAACPRI
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:24 has a nucleotide sequence of SEQ ID NO:26, as follows:
  • ccttacaggttatcacttaccgcctccgttttcgaatatttatcgttcgctagttaatcttaatttaa
    acttaaatgaggcaaataaacgtttaaactattctcttgatcgtgtgtctgattgtcttgttgtttaa
    aatgcctcctagatcgatcgtcgtagtgcaggttgttttagacaaagttgaactgcgatcagaccgag
    accggacaaccattgagcagttattttcctattcatcgtggactaactggaagatattttctgagctc
    caaaaaatatccaaaggaagggagaacgtgaaggacgaggtcggaccggacacgcctcccctcgctaa
    tcattgaggcggaggcggcggaggcgattttgggaacactcgcaggtagattttgcgtgaacttggac
    gagggtcattttcgctttggatgaatccacgaggtggtgtcactgcacgcgcacggggccctcaaacc
    gtttgaaaaccaaaccgaaggcaacaaaacgagactctcatctcatctgactctacggccagctcaag
    tgatctgctgctggtggccgacctggcggcgtgatctcgctcccgtgcccgtctcctccatccgacgc
    gtacatggcccgccatcctcatccatccgccctccagaggaccagtccagaccaataataaaagggaa
    ggtcgacgacgggctcgctccaatccggcgaaccgcgtccccgtcagcctgtcatcccgtgggcgcgc
    ggctgtcgacctgcgcatcagcttctatgattagccaggagcaataatttattactcctatttgccag
    gcgacgttcgtccaattcgacccggcaggcagcaggcagcagctgtgctcctgtgggtgggtgggtca
    tgggtgaccacatgcatcgatggagccagggccgccgtgtgcgcagccaactctacctatccccgccc
    ccgggatgggcgatggcaactatcctatcgcaacaatatcctggggtgggggctataaaacggagcgg
    cccgcgtggggcgcgcctccatcagccgcagtcgcgtcgcgtcgcgtcgcgtccagtccaatcctcgg
    agcctcacacgggcggacgagcgggagcttctcccaatctcccctgccctgccctgccctgccgccgc
    gcttagcttcgcatcttcccctcctcctcctcctccttcctcggccaagcgaggagcgaggcgcgggc
    gcgagcgcgtcgttgagatggattcggaggcggtgcagcacgggtaagcaagcaaagcaatccatgga
    tcgatccaggacacagggaggagctaggaagaggaacaatctcatgatctcattcatctgacacagcc
    ttctccctctgtctgcctgtcctcctaccgccaacagctgcgcgcattacagccgtgggtgcagcgtc
    gtggcgccctgctgcggccaggccttcggctgccgccattgccacaacgacgccaaggttcggtggtt
    tccctccttccgttttcgcttcggctccggttcagcagatgttctgaaacaaccctgtcccgtgcccc
    ggcagaactcgctggaggtcgacccgcgcgaccggcacgagatcccccgccacgaaataaagaaggtc
    agcgttccctccctctgctcaaagagcaatctcctgcctgtttcaaccattgcctatctcgtgttcgt
    ctttgttattaccgtgagcaaagaaaggaagaaacaaacaagagcgccgccttctctcttctccttct
    ccatgtaatggagcatttgttccgccgcgtagtcgagtgcaagcagcggttttcctcttttggaaacc
    cacccccacgcacggttccgttccaatctcgcccttccaattgaccaacacaaacctttcctaagatt
    tcttgtcctccttacccttctacagacaagtacgaaacgcaatcgcacaaggttatactactactagc
    ttttagtgttctagcgaccttagatttttttttttggttaggcatccctgatttttctcacacttaaa
    agcttcttcagataaggccatatcagctcagctcagtgctctgggagccgttctgcacttcacttgcg
    tgtcactaaaaacttgactgctttccgcgatgtgctccgcaccaagtggccggcactgcgtggtccac
    aggatttttcaagagaaagccggggtcacgggtgccacttgaagccaaggacaggcgtctgggattgg
    agaatatatgagaaagggataccgtcagaggcacatctcaccgtcaaactgaacagggtctaactgct
    tccagctgatttgattgagtttgagtgctgcatagttgaggacctggatatagtgacgtgtcctgaca
    ggtgtctttggacctattagcagtgaatctgacgtcgatcggctaaagcaatcatgttcgattcttat
    ccttttttttttgagagtatatgcctgattcgataaacgttcctatcctgtttcctgatgatgcatat
    atgttgtttcgattcatatagaatcataccatccatctattttgtttaaaaaaaaaatttctgggtgg
    ccatgggcacagcatgcctgttcttaagataacgattccagtaactgttcccttctgtcactgaactc
    atatgaatcgagacttaactggagctgttgcgcaggtgatctgttctctctgctccaaggaacaggac
    gtaagttgtctaccaaaacgtactcctacaacagtttttcaggagcacgcatcttttggctgtactac
    tactgctaactgcatggaaactgctcattcccatcggcaggtgcaacagaactgctccagctgtgggg
    cctgcatgggcaagtacttctgtaaagtatgcaagttcttcgatgatgatgtaagcgtactcgaatcc
    cagacgatgaacaaagaaactgaactcgatgcgttgtttactgcgtttcttttttcccccttcttctt
    cacgtacatactgtactgctcttggtccaggtctcaaagggccagtaccactgtgacggatgtggaat
    atgtaggtaagcaccaccacgctgatggctacgtctaaagacttgacgcgcagaagtgtaaaacttct
    gtcagccgttcaaaactgataaattcgggttcctcgtcttcttgttgtttatgcagaaccggcggcgt
    ggagaactttttccactgtgataaatgtggtgagtttctgcgcggacttctttctgctaagattctgt
    aaccatgctatgcagcaaagatttcactcgcgcccttattggtgtccttgttgccgtccgacagggtg
    ttgctacagcaatgtcttgaaggattcccaccactgcgtcgaaagagcaatgcatcacaactgccccg
    tctgctttgaggtgagagacctccgtttcaacggaacattcactctgaatgttccaatctcttgatat
    tgagaaggtttccctctgttttttttcacagtatctgttcgactccacgaaggacatcagcgtgctgc
    aatgtgggcataccatccatttggagtgcatgaacgagatgagagcacaccatcagtaagcatatata
    ccgttctcttcggagctgagaaacggtgccacctcacaacatcctcttttagtcgcagtgaccttaca
    gtctcagccctgtttggtctttggcagcttctcatgcccagtgtgctcgaggtccgcctgcgacatgt
    cggccacatggcggaagctcgacgaggaggtcgcggccacgccgatgcctgacatctaccagaagcac
    atggtaagaagccgaccgcccactcgttcgtcgtcccgttacatcttttccacagccatggctcgctg
    tttgacgagctctgaacctgtccggggtgccgattgctggaactgaacggcaaatgaacgctgtggtg
    tgagtgcaggtgtggatcctgtgcaacgactgcagcgcgacctcgagcgtgcggttccacgtgctggg
    gcacaagtgccccgcgtgcagctcgtacaacacccgggagacgagggctgcgtgccccaggatctgag
    gcgaaccagaggccatgtcacaaaatgccagggagatgccgtccaacgatcatctgtctgcaggacgt
    tgctgcgcttaaggttaaaggctagcgcgagaccaggcctggtagtccagtcttgagtttggtgctgg
    agcatttgtaatgttccggtaaaatgtaatgcgtccatgagtgctgtccaggcagtaagcacacctgt
    ggatcggggccggcgcaaggtccctaggcaagctgcaggattagtggggctattcatgtttagggcgc
    gaatgcaacgaaattacccgtgggccgtgggctcggtatgtaacagaaccgattatttctattacaat
    aataacatgcagttctattgggccgagcctaatcaggcaccacgaatgtgaataattgcacatggcgc
    atatatggcgggcagtagatacatataaaatggagaaaatccgtttattgccatcaaaacttatactg
    atcactacaataccatctaaaatgatgtgctcccttcaaaaccattggtttatattttctatcctttc
    attgccattgccgttacataatggcgcatgtggcatatatgggcggcgtatgtaggttcaggatggtc
    acacgacatatgacgagactacagagacatggtcaagagaagttcagtggattgtgacattctgatct
    aagacttcttaaaattggagttaataagatcgataatactcgtaccaataatgcacatcttgcttttt
    agaagcctgttttgaaataaccccaggattaagcatgtttggcccaaagaaatttttgtgatggtggt
    cgaccgagaagttttcttgactgcatgccagtgaggacaaaaaaacacatatgaaagaatcgtgttgg
    tctgtgagaatagatcaggaagttttctcgactgcgcgaccatggatggttggggtgtttcatcaagg
    atccgctttaaacacatacctttttgccttggtgatggataaggacataccggacataaaggggataa
    cccttggtgtgtgtttttttgcagacaatgtaatgctagttgatgaaagtcgggcatgagtaaatgga
    aaactagagttgcggcaagaaactttataatcaaaaggttttagacttagtagaactaaaatagaata
    tattggatgcgatttcagcactacatatgaggaatgagatcttagtttagaaggtctaggaaggacac
    ctttagatatttagtatcagcctatagagagaccgggatattaatgaagatgctagccataaaatcaa
    agtagagtcagtgaagtggagtcaagcatctggcattttatatgacaaagtgggaattgcaaaagcta
    aaaagaaagttttaggacaacgattagaccttctatattatatggaacataattttagcctacaaaaa
    gatgatatgtttagcagataaatgttgcggt&atacatatgttacgttggatttgtgaacatacaaaa
    agggatcgagtttagaatgatgatatacatgatagactaggggtagcaccagtcgatcgatatggttt
    gaatatatccaacggagacctatagaggtgtcaatatgtcttaggacctgtttgaaagcatccagttt
    ttaagaaattggtttatagaaattaaagtggttccaaacatacaagtttatgccccagtttatataaa
    ctggattatcaatttcttaaaaaccaagaagctagtctttgctagctaaaaccaacttttgcttgttt
    aattacataatgcccttgttggttgcatggaatttacatctattgtcgtcgcttttaagatagaggaa
    gggtatgttagtaattgtgtatcaaaaaatagaaaatttgtttcttagaactaagttccaaacaccct
    cacccaacttttttataaactagtttctataaactggagatagaaattggtttttaataaaccggtat
    gctttcaaacaataccttaggattctaagatgtgataccaatgagaaaaggaagaggaagactgaagt
    tggtatgggaggtgataataaaatgagtcttgaaaaaatgagatatatctaaagatttagccttgaat
    agaaatgcatgaaaataactatccatatgtttgaaccttgactttgagttttgttgaatttttaactc
    tagcctacgccaatttgtttgggactaaaaggttatgttgttgttattgccgctataaatggtgttca
    acactttcttcaagattatgatattttgttttctacaccaacaatattactgttggggtctccttctc
    tgccgaaggtcctcaggatgaagaaactgtctttggttcatcttggtaagatacgtcaaaaggaccga
    atgccgaagctgtgacagacatgcagggaatatagcagagcttcgataagagttaaagcttcggctta
    agatgattatgaaggtcatacaagaaaccaagccaccaatgaaaagacctgtttatccttaaaatttg
    tattagaacaatgtatagatatcagggtcataaatgtacttttgcttgggcggcgtcccgtgcctata
    aatagatgaactgtacccccgtactgttgacactttcattgaaagtcattctcgcactctctccttca
    agcaagacgaaggtactaatgtaatataatgtttgtaatggttcattagaatgttatccaaactatgt
    cattaccttgatatagaaaataaagtgaattcataagataataccacattgtgatattatctccatga
    gaaatgaagatccgctcttcttcaccctcgcccaaaaaccatcatctttgagagaagataattgaaag
    gaaattgggttaaccatttcctataactaattttggtgggtgatgatcaacacaaacccatggactaa
    ctagtttgtctagaattcatggattacaggtgcataaggttcaacacaaaccaagaaagaaatccggt
    tagggacacaattaaaaatggagcaaagacttga

    (>MAGI4—73717 MAGI4.contigs_w_singleton.fas 7106 bp).
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:27, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAAGCTGC
    TATTTTCTTCTTACATTGTCCACTGCGCTAGCTAGCTCGCATCTACCTGG
    AAAGCTGAAAGCTAGCCAGAGCGCTAGCTAGCTTCGTTCCTCGTCGCCGC
    GCGCCGGCCAG ATG ACTGCTCACCAGACTTGCTGCGATGATGCCGTTGC
    CGCCGGCACTGCACCGGCTGCCAGGAGGAGGCGCCTCAAATTGACGAG
    GCCGTCGGCCTCGCTCTTGATGGCGAGGAAGCTAAGGAAGAAGGCTG
    CCGGCAGCAAACGCCCAAGGGCGGCAGCGTCGAGGAAGCGCGCGAT
    GGCGATCAGGAGGAAGATGGAAGCGCTGAGGCTGCTCGTGCCACTCT
    GCGGCCGAGACAACGGCTCGGTGACCGGTGGGGCGGTCGAACGACT
    GGACGAGCTCCTCATGCACGCCGCCGGGTACATCCTGCGCCTCCAGA
    TGCAGGTCAGAGTGATGCAGCTTATGGTCCATGCACTAAATGACCGG
    CCCGAGGATTAATCTTCTTCCCAAGACCATGTGATCTTCCTTCTTTAATT
    TCTTCTTCATCTTCTTCGCGTGCCTGTGTTGCACGAGGCAGCTGTGCGTC
    GGTGTCTGGGTGCAAATCA

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW61A10-Full_Length cloned into pCR4-TOPO) (derived from MEST61-A10, GB_ACC# BM073122).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:27 has an amino acid sequence of SEQ ID NO:28, as follows:
  • MTAHQTCCDDAVAAGTAPAARRRRLKLTRPSASLLMARKLRKKAAGSKRP
    RAAASRKRAMAIRRKMEALRLLVPLCGRDNGSVTGGAVERLDELLMHAAG
    YILRLQMQVRVMQLMVHALNDRPED
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:27 has a nucleotide sequence of SEQ ID NO:29, as follows:
  • taaatctgacctccaaaatgtctctaatgaaagtgtctgcgagaaacagtattgctcgaccgaggaag
    aaaggtatttataacacacccatcacaaatggttggcttaattaaccgcaagtgcagaatagatttgc
    actgttggttcacttaataaaactgcaagtgaaaatatactttttctactatcggttttgttaagtga
    acctaccgattttgcactggtgattatttaagccaacctcatgtgataatatgaatgatttgcacgtg
    gttttattaagcgaactaacaatgtaaatgtgtttccaatggtggtttttaagtcgggaccgtcattt
    tactttaactggcgcgcaccgtgctgttttatacttgactgatgaaccttcgtggtgagtggaagcgg
    tgtggagcgagggctagctcatgcggccagccggcgacatttctcttgttccgatcccccggccggcc
    aaccactcaattaagtaggtgatcgattggcatgcatgcatggatgcatatcagcaaatgcatatcat
    atgcctcgctagctggctagtatatatagtggatgtggatcggatcatgtgacggccgggcggtggct
    gcattgcattggccctgcatatatgcacggtgacacaacaacggggcccaaataaaggacacgtcgaa
    ggtgcgcgccccagtggcgtccgacagcgcgttttgacgaggaaaagagggtgcgggcacgcgcgcac
    gcatatgctcgcggcatgcagcctcagtggccgatgacgagtggcgtgtggtgtggccggcggccggc
    cggccgggtgcctgcgtggtgcatgttgcttgccatgcatgcgtgaaatgagccgtcagcgagcgagc
    tgagggcgggcatgtggctgcatgtggccactagtttggagaacatgcggcatatgccccggaccttc
    ctgggcgctcaagcaaacaccgctctcgtgctcgctctcttgggaaatcgaagatgcatgctacccaa
    cgtgacctggatctcttttacgtacgcacaccctagcgtgctgctctcctgtgtccccgcctcctgct
    agctgttcacaatatccacgcgatttaacaaacagatatgtgtgcatgctactgcttgttttcctatt
    caatatagtaatctgctttatttagagtaccgtacctgtgccgtcagtgcccccaaccccaacgtaac
    tacgcacgcacatggcatctaatctatataagcatcagaccttgctcccttaatctcgcgctgctatt
    ttcttattacattgtccactgcgctagctagctcgcatctacctggaaagatgaaagctagccagagc
    gctagctagcttcgttcctcgtcgccgcgcgccggccagatgactgctcaccagacttgctgcgatga
    tgccgttgccgccggcactgcaccggctgccaggaggaggcgcctcaaattgacgaggccgtcggcct
    cgctcttgatggcgaggaagctaaggaagaaggctgccggcagcaaacgcccaagggcggcagcgtcg
    aggaagcgcgcgatggcgatcaggaggaagatggaagcgctgaggctgctcgtgccactctgcggccg
    agacaacggctcggtgaccggtggggcggtcgaacgactggacgagctcctcatgcacgccgccgggt
    acatcctgcgcctccagatgcaggtcagagtgatgcagcttatggtccatgcactaaatgaccggccc
    gaggattaatcttcttcccaagaccatgtgatcttccttctttaatttcttcttcatcttcttcgcgt
    gcctgtgttgcacgaggcagctgtgcgtcggtgtctgggtgcaaatcattggctgagtgtgttattgg
    tgatattatttgttcgtatatacagaatatatactcatgcatgcatactgtatgagatgatagagtaa
    atctagacatatatagttcaaggaaacctacagccaacagttgtatgcatgtgaggggggttccttgt
    ctgtatgtacgcaattgtctattgtgtgacggttgaaattgaaatttcgtcaatcatcatttcttcgt
    ctagataacgtgtgtacaaacggcgagtgtttaaatgaactagagctaataattagtggctaaaatta
    gctggagacatccaaacaccctaactaataatttaactattagttatttttagtaaattagtcaatac
    ttagctagctatttgttagctagctaattctactagcattttttagctaactagctattagctctagt
    acattcaaacacccttttagggactaatttttagtctctccattttatttcattttagtcactaaatt
    accaaatacgaaaattaaagctctattttagtttccggtatttgacaatttag

    (>MAGI4—145622 MAGI4.contigs_w_singleton.fas 2433 bp).
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:30, as follows:
  • GGACACTGACATGGACTGAAGGAGTAGAAAATCCATCCATTCCCCTCGCC
    AAGCCGCCACGGCCTGACTTTCCCTCCCGCACACCCGCGACCATACAGGC
    AAGTCAGGCATACACCAACAACGCTCGTCGTGCACCTCGCGCCTCAGGTC
    ACCCCACCAAATTCCTCTTGATACGCCGAATTTCTTTTGCTAATTCTGCT
    ACCTCCTGTCGCTAAGCCACCATATTCAGTCTAACCCCTGCTCTGAGCTC
    ACCTGATTGGCGGCTCCGTTCGGCCTCTGGGCCTGGGTGTACCGACTACC
    GAGGGCTCTTTCGAAATGTCAATTGGGTCGAGTTTGGTGGGCTACGTGAA
    GC ATG GATGAATTTCCCGGCTGGAAGCGGGAGGCGGCAGCAGCATCCGG
    GGCCGGAGCACCTGTCGCCGATGACGCCGCTCCCGCTGGCGCGGTAG
    GGGTCGGTCTACTCGCTCACGTTCGACGAGTTCCAGAGCTCGCTCGG
    TGGGGCCACCAAGGACTTCGGATCCATGAACATGGACGAGCTCCTCC
    GCAACATCTGGTCGGCGGAGGAGACACACAGCGTCACAGCTGCGGAC
    CATGCCGCGCGGGCGCCGTACGTCCAGTCCCAGGGCTCGCTCACCCT
    CCCCTGCACGCTCAGCCAGAAGACCGTCGACGAGGTCTAGCGTGACC
    TCGTGTGCAACGGTGGAGGACCCTCCGACGAGGCTGTGGCGCCGCCCCAC
    CGGCCCAACGGCAGCCGACGCTCGGGGAGATC ATG CTGGAGGAGTTCCT
    CGTCCGCGCCGGCGTGGTGAGGGAGGACATGATGGCGGCGGCGCCC
    GTACCACCAGCGCCGGGTTGCCCACCACCTCATCTGCAACCGCCAAT
    GCTGTTTCCACATGGCAATGTGTTTGCTCCCTTAGTGCCTCCGCTCCA
    ATTCGGGAATGGGTTTGTGTCGGGGGCTCTCAGTCAGCAGCAGGGAG
    GTGTTCTTGAGGCCCCGGCGGTATCGCCGCGGCCGGTGACGGCAAGC
    GGGTTCGGGAAGATGGAAGGAGACGACTTGTCGCATCTGTCGCCATC
    ACCGGTGTCGTACGTTTTTTTGTGCTGGTTTGAGGGGAAGGAAGCCA
    CCAGCTGTGGACAAGGTGGTTGAGAGGAGGCAACGCC

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon, coding sequence in bold) (Sequence of 5′ RACE product CW76H12-Full_Length cloned into pCR4-TOPO) (derived from MEST76-H12, GB_ACC# BM073865).
  • A predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:30 has an amino acid sequence of SEQ ID NO:31, as follows:
  • MDEFPGWKREAAAASGAGAPVADDAAPAGAVGVGLLAHVRRVPELARWG
    HQGLRIHEHGRAPPQHLVGGGDTQRHSCGPCRAGAVRPVPGLAHPPLHAQ
    PEDRRRGLA
  • Another predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:30 has an amino acid sequence of SEQ ID NO:32, as follows:
  • MLEEFLVRAGVVREDMMAAAPVPPAPGCPPPHLQPPMLFPHGNVFAPLVP
    PLQFGNGFVSGALSQQQGGVLEAPAVSPRPVTASGFGKMEGDDLSHLSPS
    PVSYVFLCWFEGKEATSCGQGG
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:30 has a nucleotide sequence of SEQ ID NO:33, as follows:
  • tagaatagccagcatcgacaaattacttacaaatagaaacattacctgtttcctcccacgcgacctcg
    cggccaactcccggttcttgatcatccggcgttgcctcctctcaaccaccttctccacagctggtggc
    ttccttcccctcaaaccagcacaaaaaaacgtacgacaccggtgatggcgacagatgcgacaagtcgt
    ctccttccatcttcccgaacccgcttgccgtcaccggccgcggcgataccgccggggcctcaagaaca
    cctccctgctgctgactgagagcccccgacacaaacccattcccgaattggagcggaggcactaaggg
    agcaaacacattgccatgtggaaacagcattggcggttgcagatgaggtggtgggcaacccggcgctg
    gtggtacgggcgccgccgccatcatgtcctccctcaccacgccggcgcggacgaggaactcctccagc
    atgatctccccgagcgtcggctgccgttgggccggtggggcggcgccacagcctcgtcggagggtcct
    ccaccgttgcacacgaggtcacgctagacctcgtcgacggtattctggctgagcgtgcaggggagggt
    gagcgagccctggcactggacgtacggcgcccgcgcggcatggtccgcagctgtgacgctgtgtgtct
    cctccgccgaccagatgttgcggaggagctcgtccatgttcatggatccgaagtccttggtggcccca
    ccgagcgagctctggaactcgtcgaacgtgagcgagtagaccgacccctaccgcgccagcgggagcgg
    cgtcatcggcgacaggtgctccggccccggatgctgctgccgcctcccgcttccagccgggaaattca
    tccatgcttcacgtagcccaccaaactcgacccaattgacatttcgaaagagccctcggtagtcggta
    cacccaggcccagaggccgaacggagccgccaatcaggtgagctcagagcaggggttagactgaatat
    ggtggcttagcgacaggaggtagcagaattagcaaaagaaattcggcgtatcaagaggaatttggtgg
    ggtgacctgaggcgcgaggtgcacgacgagcgttgttggtgtatgcctgacttgcctgtatggtcgcg
    ggtgtgcgggagggaaagtcaggccgtggcggcttggcgaggggaatggatggatatgtgtcgccacc
    aaggagtcgtgtgggggagtttaaaacgtcgcaaggctcgaggtcgcacatggtgttgggtttgggtg
    cgtgctgggtcataaaagctgaaagggaattaggcttacacctatttcctaaatgattttggtggttg
    aattgtccaacacaaa

    (>MAGI4—7232 MAGI4.contigs_w_singleton.fas 1376 bp)
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:34, as follows:
  • ATTCCCGTCTTACCTAGCGCTAGGGTTAGTACGCGTCCACGGCGACGACC
    TCTGCGCGGAGTGTGCTCCGATTGGCTGGCCTCCTCGATCCTCCTTCCCG
    CGAACGCACGCGCGCGCGAGGGAGAGGTTGAGACTTGAGAGATAGACGAA
    AGACGAAACAAGGGAAGGAGACGCCGTGCTCGCCTATTGGCCGCCGCCTC
    CGCTCCTTCGCGCCCAATGGCTTCTGCAGCATATCAATATC ATG CAGCAT
    AGCAGTACTCAGACCCTTACTACGCAGGCGTTGTTGCTCCCTATGGAAG
    TCAAGATGTGTGTCCGAGGAGCCTGTCTATGTGAACGCCAAGCAGTAC
    CGCGGCATTCTAAGACGGCGGCAGTCACGTGCCAAGGCCGAGCT TGA
    GAGAAAGCGCTGGTCAAGCAAGAAAGCCGTATCTTCACGAGTCCCCGTCA
    TCAGCACGCGATGACGAGGAGGGCGAGAGGGAACGGTGGACGCTTCCTAA
    ACACGAAGAAGAGTGACCGTGTCCCTCCTGATGACTTGATACAGCTACGA
    CGACACAACGAGGCTTGAAGAGGTAGCGGTCTGGCTGGCATCCTAGAGCA
    GCGGTTTCTGTCCACAGGCACGTGCATCTGAGACCGGATCCGTAGCTCCA
    CTCCACAGCATATGCGCAGCCCATCCATCTCGTGCACACTTG

    (Underlined=start and stop codons; coding sequence in bold) (Sequence of 5′ RACE product AM77A01-5T3 Full_Length cloned into pCR4-TOPO).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:34 has an amino acid sequence of SEQ ID NO:35, as follows:
  • MQHSSTQTLTTQALLLPMEVKMCVRGACLCERQAVPRHSKTAAVTCQGRA

    (The above sequences are presented after trimming GeneRacer Oligo sequence. Cloned in pCR4-TOPO vector at the “TOPO Cloning site”.)
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:36, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAAAAAAA
    CCCAAATCAAATTTCGCCTTCGTCGTCGTCTTATCGTCTCAGATTTGACT
    CC ATG TCGGCGGCGCTCGCGGTGACGGACGAGGTGGCCCTGCCGATCC
    GGGCGGTGGGGGATCTAGCGGCCGCCGCCGAGGTCTCGCGGGAGGA
    GGTCGCCGTCATCACCCAGTGCGCGGCGCTCGGTGGGAAGTTGCCTT
    TTGAAGATGCATCAGTTGGTGCGGTTCTTGCAGTCATTAAAAACGTGG
    AAAGCTTGAGGGAGCAATTGGTTGCTGAAATCAGGCGGGTGCTGAAA
    GCTGGTGGAAGAGTATTGGTGCAGAGCCCTGCACCCTCATCCAGTCA
    GAAGCCGAACACTGATATTGAGCGCAAGTTACTGATGGGTGGATTTG
    CTGAAGTGCAATCTTCTGCTGCAAGCTCGCAGGATAGCGTGCAATCT
    GTTACAGTTAAGGCAAAGAAGGCTAGCTGGAGCATGGGCTCTTCTTT
    TCCCCTTAAGAAAACAACAAAAGCCCTTCCCAAGATTCAAATTGACGA
    CGACTCTGATCTGATTGATGAAGACAGTCTCTTGACTGAGGAGGACC
    TGAAGAAACCACAACTTCCAGTTGTTGGGGACTGTGAGGTGGGGGCA
    GCAAAGAAAGCATGCAAGAACTGTACTTGTGGCAGGGCTGAGGCCGA
    GGAGAAGGTTGGGAAGCTGGAGCTCACTGCGGAGCAGATCAATAACC
    CTCAGTCAGCTTGTGGCAGTTGTGGGTTGGGTGATGCCTTCCGCTGT
    GGAACCTGTCCCTACAGAGGTCTTCCACCATTCAAGCCTGGCGAGAA
    GGTTTCCTTGTCTGGCAACTTCCTTGCTGCTGACATATGATGGCATCG
    CCAACATCGGCAAAACAAGGA

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ ACE product CW88H03-Full_Length cloned into pCR4-TOPO) (Derived from MEST88-H03, GB_ACC# BM079064).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:36 has an amino acid sequence of SEQ ID NO:37, as follows:
  • MSAALAVTDEVALPIRAVGDLAAAAEVSREEVAVITQCAALGGKLPFEDA
    SVGAVLAVIKNVESLREQLVAEIRRVLKAGGRVLVQSPAPSSSQKPNTDI
    ERKLLMGGFAEVQSSAASSQDSVQSVTVKAKKASWSMGSSFPLKKTTKAL
    PKIQIDDDSDLIDEDSLLTEEDLKKPQLPVVGDCEVGAAKKACKNCTCGR
    AEAEEKVGKLELTAEQINNPQSACGSCGLGDAFRCGTCPYRGLPPFKPGE
    KVSLSGNFLAADI]
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:36 has a nucleotide sequence of SEQ ID NO:38, as follows:
  • gctgtaccagttgaggtactccttgacgtcctcgtacatggtgggcgccagcgggtgccagatgccgg
    agtcgaggtagagcacggggtcgtcgtacttgatgccggcgcccctaagcaccggcaagtaggatccg
    gcgatcatcttgaggaagttctggaggttgtccgccgagccgccgagccagaactggaggctgaggat
    gtacagccaggcgtcctgcgccttgtcggagggcaggtacttgagcaccttgggcagcgtgcgcacga
    gcttgagcatgctgtcggcgaagttgctggagttggacttgctgcgcttgaagagctggaagaagggg
    ctcttggactgccccagctgcgacatgctgaaggagccgagcttgttgaggcgcatgacctcgggcat
    ggaggggaagacaaggacggcgtccatgcggtcgcgctccttctcggccgcggccttgaccttgagcg
    ccagctcatcgacgaagatgagggagccgatgaagacgttgacgtcggcgaggtcggcgaggaaggta
    gcccacgacctcgaacgacgcggcgcggcgcgggtcagcgttgagctgctgcacggcggccgtgacgg
    acgactggtactgcgcctccagcacgacgtagacgaccttgacccgaggcaggccccgcgggtcggcc
    ggcaccacgcgccgcacctcgggcttggtctgcgtgaacaagccgttgccgccggcgaccgcgcaccg
    gatggcgccggcgcgccgcggctgctggcgacggctgctcaggaggaacgagtgcagcggcacgggcg
    ccgccaggagctgcttctgcgccgcggaggcgaatggggtggacactagcgacgacgacatggcgcct
    gctcacaggacggagccggcgggcggagaaacgcgcgcctggacactgacgcgacgctcgagcgcagt
    aagtaaaaaaaaatctacactagactactagagtaaggcgcctgttcttggctcgtggctggacaatt
    gttcttggcggccgccgtccctcggaaacagagcagggaaaggagaagaagcgagcaggggagcgcgg
    gaggcgggaaaatgtataggttgtccgtgtccacgtccttcgtctcaattaagaagaggcatccaggc
    tcacaaaatcaatctgaaaacacatgcactgatgcacacttgtgtttgtgtagaggcgcttatatatc
    atccaaaagacaagtcactcacacgcaaattcgcattggctaacagaagctatttggaatgcagttca
    gtcgactaacaacgtaggtacccccgtctccttgttttgccgatgttggcgatgccatcatatgtcag
    cagcaaggaagttgccagacaaggaaacctgccaatcggagaagcagcagcagtgaacgttcaagatc
    cagagtacaatcgacagacatattttgatctcctcgagaattctatcaggggaggagacgagtagaac
    tgttttaccttctcgccaggcttgaatggtggaagacctctgtagggacaggttccacagcggaaggc
    atcacccaacccacactgcaaagaaaaatcaaggatcatttacagatatcaccagacgtgataggtaa
    cctagtccgagtgaacgtatgaaatttcacgagggggcacaagtgccacctgtaagcaatacttacac
    tgccacaagctgactgagggttattgatctgctccgcagtgagctccagcttcccaaccttctcctcg
    gcctcagccctgccacaagtacagttcttgcatgctttctttgctgcccccacctcacagtccccaac
    tggtgaaaacatcagtgaaaacatcacttaactgtttaggatccaaacctaaactggctattgcttac
    ggagttgaactaagttgacgggttttgttgctctaccaactggaagttgtggtttcttcaggtcctcc
    tcagtcaagagactgtcttcatcaatcagatcagagtcgtcgtcaatttgaatcttgggaagggcttt
    tgttgttttcttaaggggaaaagaagagcccatgctccagctagccttctttgccttaacctacaagt
    ggttcaaattagcacaaaaactaaagcctgcacagcaaaactaacatactataacacatgatcttaga
    ccactcactgtaacagattgcacgctatcctgcgagcttgcagcagaagattgcacttcagcaaatcc
    acccatcagtaacttgcgctcaatatcagtgttcggctaacggagacaatcataaaaaaattaagaac
    tttaaatcgacattgcaagagaaacgagacaacaaagacagattctgataagttaataccttctgact
    ggatgagggtgcagggctctgcaccaatactcttccaccagctttcagcacccgcctgatttcagcaa
    ccaattgctccctcaagctttccacgtttttaatgactgcaagaaccgcaccaactgatgcatcttca
    aaaggcaacttcacacctgatgcatggcagaacaatagtttggtcacggttttgtgataacacacaca
    cacacacacacacacacacacacacacacacacacacacacacggcatagcactagcaaagcataaca
    caaattaaaaatcgaacattattgtttaatagaggctcccaaaatcaggaatgctagcacttggctta
    ttcataaacacacacatccataatcaggaagcatacattactgaaccattaaatttaataataaaaat
    tcagatgttgaatccatggctgaaattttctgttccttttgaaagtataatcctaactttcatctccg
    gctgacctggtaatatcttctagctccttttaccttatatttttttcagttgcttgagaaatagcggt
    aggaaaattgacacatgtcattcgtaaatccatgggacttagagcaactccaagagcttcctaagaaa
    ttgttccccaaaacatcatatagggggctgctgaaaaaaatccactaagagcaactccaaatgagtgc
    tagaaaatttccccaaaaaatgattattggggatatgttaaaaaattttaggggtgaattatcatgta
    tactccaacgattccgttaaacaaatgcgactcaatctcagccacagtctgagtcttacagacacaca
    caaaacctaacatgccggtggcagccacattatcacacaccggaacaaataactttgaggcaaaaaca
    cattatgcaagcagagaaacaccagaacagactcccagctgttgaagtgcaaatgtgttttctatatt
    tgagttacttgctggtaaatcccgatcgggaatgtaataatcggggagttgcattagcacttttgcag
    caagctaagccaactggttgggaatgtcaagcattcttgagcaggagtactagtcaagttaacaggct
    tcagatcccatccaatcattgtcacatttgaatataacttgagcgggtagaaaaaatatcataacaaa
    ggcatcatggactgaatcctaaacatcataacgaaggcatcatggactgaatagcgatcatcataaca
    acggcaggaaacagactcccaactgaatcatggttaacatggactgaattgtggtggcactgcatgca
    gtgcgagatgcatcatatccaggtcaattcaggttagcaaatgcaaggccacaggagttgccgccagg
    gaggaggctctaggcgaggtcacgggagttgcggtggaagttgctgcggattggggaagacctttgct
    cgccaatatttgagggagagtggagctcggatgcgggacgctgataatttgggggaaggaaaggggaa
    ctattgggtggagaattttttgtttttcaccccaaaacatgtttttgggttggttttagcgttcttct
    ggagatgctcttaagcaactagcacatgagacatggcatagatatcaagaactgcaaggagaggttca
    agttcaaatctgaagaagtctgcaagggcatgtccacagattcagcggttttggagttgggaaataac
    ttcagctttcttttctttttgttgttgagacgttcttttctttttcttttttttttgttgttgttgag
    gcgtcagctcgacgttttcattctacacattagaaagtggcagtagcgcaagagataccacagggcca
    aaactactagtggtactgaaagttttcattcgaagaatcagtaagtggcactatcacaggaagaaaca
    ttgcaaggccaaacttggcgtccactgactgcgcttcaatattacttgagcaacttgctagcctcccg
    atcccggaaggatggtttgataaactaattctctaattgaagtgggaacccttaagaaccaaacgtcc
    actactccaaatttgattgcaaaagaaaaaagaatctagcccattccgcggaatcacgccagaaggct
    cgctaattgaagcatgcaagcaaggcagcaaagagaacagcacgcatcgacgggttcctgcatccaca
    agcacgaacttggcaacttgccatggtcgcctcgagggaaagaaatagaagaaaaaatggaaagaggg
    caagacgggggcgaaaccagctaagctcaccgagcgccgcgcactgggtgatgacggcgacctcctcc
    cgcgagacctcggcggcggccgctagatcccccaccgcccggatcggcagggccacctcgtccgtcac
    cgcgagcgccgccgacatggagtcaaatctgcacacgagcacacgccgagaaccagaagagactcggt
    gaaaggagtatccccgaagagaaaaggaattagggttaatcgagggagggttttatctgcacgccccc
    ggattcatcacgcgactgctacctgagacgataagacgacgacgaaggcgaaatttgatttgggtttt
    gcctggcctcctctcctctcgaagcttcacaacacgccgagttatttgatattgtaacaatctcgtcg
    cgcggcttcaccagttattactccgtagttatacttcgctagtttagtatt

    (>MAGI4—101388 MAGI4.contigs_w_singleton.fas 5083 bp).
  • A suitable nucleic acid molecule that is modulated (e.g., up-regulated) by nitrogen is the non-symbiotic hemoglobin gene (MEST129-C09.T3Seq) from corn having the nucleotide sequence of SEQ ID NO:39, as follows:
  • catccatccatccatccatttccaatcccaatcccaatcccaccagtgtccagtgctcggggaaccgac
    acagctcctcagcagagtagccagcacgacaagcccgatcagcagacagcaggcatggcactcgcggag
    gccgacgacggcgcggtggtcttcggcgaggagcaggaggcgctggtgctcaagtcgtgggccgtcatg
    aagaaggacgccgccaacctgggcctccgcttctttctcaaggtcttcgagatcgcgccgtcggcgaag
    cagatgttctcgttcctgcgcgactccgacgtgccgctggagaagaaccccaagctcaagacgcacgcc
    atgtccgtcttcgtcatgacctgcgaggcggcggcgcagcttcgcaaggccgggaaggtcaccgtgagg
    gagaccacgctcaagaggctgggcgccacgcacttgaggtacggcgtcgcagatggacacttcgaggtg
    acggggttcgcgctgcttgagacgatcaaggaggcgctccccgctgacatgtggagcctcgagatgaag
    aaagcctgggccgaggcctacagccagctggtggcggccatcaagcgggagatgaagcccgatgcctag
    tagtggcgattgcgaccagtgtttaacccatgacgcagcgccgtcacagatgtcccgtgtggtcttgcg
    ctttagcaatttctctctggagggagcgtgtattgttatcttgtgatcgagagcctgtgtgctgccttt
    gcttcttgtgattatatagctactgaataaagatgtagcgttcttcaaaaaaaaaaaaaa
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:39 has an amino acid sequence of SEQ ID NO:40, as follows:
  • MALAEADDGAVVFGEEQEALVLKSWAVMKKDAANLGLRFFLKVFEIAPSA
    KQMFSFLRDSDVPLEKNPKLKTHAMSVFVMTCEAAAQLRKAGKVTVRETT
    LKRLGATHLRYGVADGHFEVTGFALLETIKEALPADMWSLEMKKAWAEAY
    SQLVAAIKREMKPDA
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:42, as follows:
  • TCGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAAATCA
    CCTAGCTAGAAAGGAGAGCACCGAGCGCTGCACCACTACTGCTGATATGA
    GCACCTGAACCTTCTGGGCAACCACATCCGGTCCCTGCCCCTGATCATCC
    GCAGCAGCC ATG GCGCAGCAGCAGGAGAAGAAGCAGCAGCAGAGGGG
    GAAGCTGCAGAGGGTGCTAAGGGAGCAGAAGGCTCGGCTCTACATCA
    TCCGCCGATGCGTCGTCATGCTCCTCTGCTGGAGTGAC TGA TCCATCT
    CAAGCATGCATGATAAACCTGTGCTCTTTTTTTTTCCTTCTGTTTTTTCC
    CCTCTTTTTCCCATCCTTTTCACCTTGCCACTTTGGTGGGCGA

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start and stop codons; coding sequence in bold) (Sequence of 5′ RACE product MEST213-C11-Full_Length cloned into pCR4-TOPO).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:42 has an amino acid sequence of SEQ ID NO:43, as follows:
  • MAQQQEKKQQQRGKLQRVLREQKARLYIIRRCVVMLLCWSD

    (The above sequence is presented after trimming. Cloned in pCR4-TOPO vector at the “TOPO Cloning site”.)
  • A suitable nucleic acid molecule of the present invention is a gene that is up-regulated by nitrogen and contained in a full-length cDNA clone having the nucleotide sequence of SEQ ID NO:44, as follows:
  • CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAAAACTA
    ACACTTCACGTGCCCCCATCCTTTTCCGCCTCAAGTCAAGTGTTCACGGT
    CCATCCTCTCGAGAGTCTAGGCCCTTCTCCCGAAGCCGCAGACGCAGAAA
    ACGGCTCTGCAT ATG GAGGCGAAGAAGAAGCCGTCGGCCCCCGCCGCC
    GTCGGAGCCGCGCCGCCGCCGCCGGGTAACGGGTACTTCAGCACCGT
    CTTCTCCGCGCCGACTGCGGGAAGCGCAAGTGACGCAAAGCATGCGG
    ACTTGTACACGATGCTGAACAAGCAGAGCTCCAGAGGGCAGAATGGC
    AGAGATGGCAAATCCCACAGCCGCCCTACTTACAAGGATGGCAAACA
    TGCTCATCCAAATGAGCCATCAGAATCTCCTTACTTTGGCTCATCCGT
    GCATTACGGTGGTCGGGAGTTCTACAGCAGCGTTTTACGGAAGCAAC
    CAGCCAATGAACCCCATACGGATTACAAGGGGGACAACCCGGATGGC
    TCTGCTACCAGAGGTGATTGGTGGCAAGGTTCACTTTATTACTGAATA
    ATCTGCTGGGACCTCTCCCTTTTGTGAACAAGGAATAAAAGGGGTAGAGC
    TGAGAATGGTTTGTTGTAGTGTTGGAAGTGTTGACGCGAGCCGTCAAGCA
    TCGATCAATAGTAATAGTTGTAATAGTTGAAAGCTGCGTCGTGACTACAA
    GCATCCTGTTGGTGGAGGCAGTATTTTAGATCCATCATCACGCCTGGACA
    GATGTGGGTGTCC

    (Underlined=GeneRacer Oligo sequence; Bold/Underlined=start codon; coding sequence in bold) (Sequence of 5′ RACE product CW264H08-Full_Length cloned into pCR4-TOPO) (derived from MEST264-H08 GB_ACC# BM350368).
  • The predicted protein or polypeptide encoded by the full-length cDNA clone of SEQ ID NO:44 has an amino acid sequence of SEQ ID NO:45, as follows:
  • MEAKKKPSAPAAVGAAPPPPGNGYFSTVFSAPTAGSASDAKHADLYTMLN
    KQSSRGQNGRDGKSHSRPTYKDGKHAHPNEPSESPYFGSSVHYGGREFYS
    SVLRKQPANEPHTDYKGDNPDGSATRGDWWQGSLYY
  • A putative promoter (upstream of the transcription site of the gene) for the gene of the full-length cDNA clone of SEQ ID NO:44 has a nucleotide sequence of SEQ ID NO:46, as follows:
  • aaagcttacacttcataagagattcatagttttatcttacagccatcgttgtcaacctcaactaccat
    gcaatccgtttgggattcaactagcaagtaaggggatgtttgtttgggtttataatctgtctggatta
    tataatctaacaacttttgaactaacacttagttcaagaattgttggattatataatctaggcagatt
    ataatcccaaacaaacacttcctaagtcttgtacaggctatagagattatttttccagaatggaggag
    ggataatgacaagacctaaaagaaagttatgtttatggaaaacaaaaaaatggagccaggataatgac
    acaaaagaaaggtatgttttctggaataaaaaaaattaaatatatattttgaacttcctaagactgga
    acatgatacctaagctggacagatgatcaaggacagttttacccctggagacagaaaaacttataaga
    cttagctttctacatcatatcctgttttgtatgtctcataattaggttccttgtattaagacgaccaa
    cctatcatttgttatacaaaattcgaacgactgctgaagtctcgaagtatatagtctaggctgattaa
    aatgtaagtatgggttaaagtgctgctggtaacaaactaaatacaactgtatgatgttgttgacaaca
    agacataactcaaaatgggagcaccaacaaagtgactggcaccggtgatgcaagcataacctaaacac
    aactaatggaaaacgcgaattggaaactatgaaagtgtcccatatatggtataccttgttcacaaaag
    ggagaggtcccagcagattattcagtaataaagtgaacctgaaagtgaagtctagcaagtcagtgtat
    gagcgtccatgtatatactgaagataatacacaaattgatgcaatgataccttgccaccaatcacctc
    tggtagcagagccatccgggttgtcccccttgtactggatttaaaattcaaaataaacattagactta
    agcgctccaaatgatctgtactacgtatatataaaaaggttctacgtacatccgtatggggttcattg
    gctggttgcttccgtaaaacgctgctgtagaactcccgaccaccgtaatgcacggatgagccaaagta
    aggagattctgatggctcatttggatgagcatgtttgccatccttgtaagtagggcggctgtgggatt
    tgccatctgagcacgaatttaaacttccatagttaaaatcagtgctccagattaattctaagctaaga
    tggtgagaaaaggttttaagtatcgttgtgcttatgaacgcgacctaaatcgaagagaaacgtcaaat
    tgacaagagtacccagaactacctctgccattctgccctctggagctctgcttgttcagcatcgtgta
    caagtccgcatgctttgcgtcacttgcgcttcccttgaatgcaaaacaaagtcaaaatgtcaacgtca
    tatccaaatagattttgcataatcctataggtcctctattatcaaaatcacccctcatcagaattaaa
    ttgggaaaccgttgaagtccctccacaaatcgcaacatagtaacggactctttcatcaaatcgcacca
    gctcactaatcatgcaaaaaaattactaagaccccaggaatctgagagcaaaatatcagaacgatggc
    gtgaagagacggcccgtaccgcagtcggcgcggagaagacggtgctgaagtacccgttacccggcggc
    ggcggcgcggctccgacggcggcgggggccgacggcttcttcttcgcctccatatgcagagccgtttt
    ctgcgtctgcggcttcgggagaagggcctagactctcgagaggatggaccgtgaacacttgacttgag
    gcggaaaaggatgggggcacgtgaagtgttagttgtaggcggcggcggccggcggggaaggaagcagt
    tggttgttcgcctcgtggcgtcctgcttcggccaacatctgtgccggcatttaaaggcctcgacggag
    cgactcggtttcgctatttcggagatcttaaggggctgaatggagaaaattgtgtttagctttcatcc
    acatccatccaacctgcagtgagacttgcagagtgcagactcccgtattacagggacggtcctgaata
    agttagtagttttatttcagagattcaacgatgttagtatacgaattatttagacacgtttggaatca
    tccagttttttagcaatctgatttataaaaagtcaagtgcttccaaacatatcagattatgcttcggt
    tcttaaaaatcggactgcctcttccataactaaaattagtttttaacttggtagaaattagtgattgt
    aaccgctcttaggtctatgcatgtgattccctcgatgtctttatcccatttgaatatttaattattat
    ttaaaaattttagattaaaaatattaattcaatctatatttaaaattggcaacaaagaaaaacaaaga
    gaataatagaatcaattacttttggaatagagtaaggattgaatttgtctttgtgtataacaaagcta
    gaagttggtttccaagaactagcctctaacacgcacacctatttttt

    (>MAGI4—139395 MAGI4.contigs_w_singleton.fas 2631 bp).
  • The present invention relates to a nucleic acid construct having a nucleic acid molecule that is modulated by nitrogen in corn. The construct also includes a 5′ DNA promoter sequence and a 3′ terminator sequence. The nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule.
  • The nucleic acid molecules of the present invention may be inserted into any of the many available expression vectors and cell systems using reagents that are well known in the art. Suitable vectors include, but are not limited to, the following viral vectors such as lambda vector system gt11, gt WES.tB, Charon 4, and plasmid vectors such as pG-Cha, p35S-Cha, pBR322, pBR325, pACYC177, pACYC1084, pUC8, pUC9, pUC18, pUC19, pLG339, pR290, pKC37, pKC101, SV40, pBluescript II SK± or KS± (see “Stratagene Cloning Systems” Catalog (1993) from Stratagene, La Jolla, Calif., which is hereby incorporated by reference in its entirety), pQE, pIH821, pGEX, pET series (see Studier et. al., “Use of T7 RNA Polymerase to Direct Expression of Cloned Genes,” Gene Expression Technology vol. 185 (1990), which is hereby incorporated by reference in its entirety), and any derivatives thereof. Recombinant molecules can be introduced into cells via transformation, particularly transduction, conjugation, mobilization, or electroporation. The DNA sequences are cloned into the vector using standard cloning procedures in the art, as described by Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor, N.Y.: Cold Spring Harbor Press (1989), and Ausubel et al., Current Protocols in Molecular Biology, New York, N.Y.; John Wiley & Sons (1989), which are hereby incorporated by reference in their entirety.
  • In preparing a nucleic acid vector for expression, the various nucleic acid molecule sequences may normally be inserted or substituted into a bacterial plasmid. Any convenient plasmid may be employed, which will be characterized by having a bacterial replication system, a marker which allows for selection in a bacterium, and generally one or more unique, conveniently located restriction sites. Numerous plasmids, referred to as transformation vectors, are available for plant transformation. The selection of a vector will depend on the preferred transformation technique and target species for transformation. A variety of vectors are available for stable transformation using Agrobacterium tumefaciens, a soilborne bacterium that causes crown gall. Crown gall are characterized by tumors or galls that develop on the lower stem and main roots of the infected plant. These tumors are due to the transfer and incorporation of part of the bacterium plasmid DNA into the plant chromosomal DNA. This transfer DNA (T-DNA) is expressed along with the normal genes of the plant cell. The plasmid DNA, pTi, or Ti-DNA, for “tumor inducing plasmid,” contains the vir genes necessary for movement of the T-DNA into the plant. The T-DNA carries genes that encode proteins involved in the biosynthesis of plant regulatory factors, and bacterial nutrients (opines). The T-DNA is delimited by two 25 bp imperfect direct repeat sequences called the “border sequences.” By removing the oncogene and opine genes, and replacing them with a gene of interest, it is possible to transfer foreign DNA into the plant without the formation of tumors or the multiplication of Agrobacterium tumefaciens (Fraley et al., “Expression of Bacterial Genes in Plant Cells,” Proc. Nat'l Acad. Sci. 80:4803-4807 (1983), which is hereby incorporated by reference in its entirety).
  • Further improvement of this technique led to the development of the binary vector system (Bevan, “Binary Agrobacterium Vectors for Plant Transformation,” Nucleic Acids Res. 12:8711-8721 (1984), which is hereby incorporated by reference in its entirety). In this system, all the T-DNA sequences (including the borders) are removed from the pTi, and a second vector containing T-DNA is introduced into Agrobacterium tumefaciens. This second vector has the advantage of being replicable in E. coli as well as A. tumefaciens, and contains a multiclonal site that facilitates the cloning of a transgene. An example of a commonly-used vector is pBin19 (Frisch et al., “Complete Sequence of the Binary Vector Bin 19,” Plant Molec. Biol 27:405-409 (1995), which is hereby incorporated by reference in its entirety). Any appropriate vectors now known or later described for genetic transformation are suitable for use with the present invention.
  • U.S. Pat. No. 4,237,224 to Cohen and Boyer, which is hereby incorporated by reference in its entirety, describes the production of expression systems in the form of recombinant plasmids using restriction enzyme cleavage and ligation with DNA ligase. These recombinant plasmids are then introduced by means of transformation and replicated in unicellular cultures including prokaryotic organisms and eukaryotic cells grown in tissue culture.
  • Certain “control elements” or “regulatory sequences” are also incorporated into the vector-construct. These include non-translated regions of the vector, promoters, and 5′ and 3′ untranslated regions which interact with host cellular proteins to carry out transcription and translation. Such elements may vary in their strength and specificity. Depending on the vector system and host utilized, any number of suitable transcription and translation elements, including constitutive and inducible promoters, may be used. Tissue-specific and organ-specific promoters can also be used.
  • A constitutive promoter is a promoter that directs expression of a gene throughout the development and life of an organism. Examples of some constitutive promoters that are widely used for inducing expression of transgenes include the nopaline synthase (“NOS”) gene promoter from Agrobacterium tumefaciens (U.S. Pat. No. 5,034,322 to Rogers et al., which is hereby incorporated by reference in its entirety), the cauliflower mosaic virus (“CaMY”) 35S and 19S promoters (U.S. Pat. No. 5,352,605 to Fraley et al., which is hereby incorporated by reference in its entirety), those derived from any of the several actin genes, which are known to be expressed in most cells types (U.S. Pat. No. 6,002,068 to Privalle et al., which is hereby incorporated by reference in its entirety), and the ubiquitin promoter, which is a gene product known to accumulate in many cell types.
  • An inducible promoter is a promoter that is capable of directly or indirectly activating transcription of one or more DNA sequences or genes in response to an inducer. In the absence of an inducer, the DNA sequences or genes will not be transcribed. The inducer can be a nutrient (e.g., nitrogen, including nitrogen in the form of nitrate), a chemical agent, such as a metabolite, growth regulator, herbicide, or phenolic compound, or a physiological stress directly imposed upon the plant such as cold, heat, salt, toxins, or through the action of a pathogen or disease agent such as a virus or fungus. A plant cell containing an inducible promoter may be exposed to an inducer by externally applying the inducer to the cell or plant such as by spraying, watering, heating, or by exposure to the operative pathogen. An example of an appropriate inducible promoter is a glucocorticoid-inducible promoter (Schena et al., “A Steroid-Inducible Gene Expression System for Plant Cells,” Proc. Natl. Acad. Sci. 88:10421-5 (1991), which is hereby incorporated by reference in its entirety). Expression of the transgene-encoded protein is induced in the transformed plants when the transgenic plants are brought into contact with nanomolar concentrations of a glucocorticoid, or by contact with dexamethasone, a glucocorticoid analog (see Schena et al., “A Steroid-Inducible Gene Expression System for Plant Cells,” Proc. Natl. Acad. Sci. USA 88:10421-5 (1991); Aoyama et al., “A Glucocorticoid-Mediated Transcriptional Induction System in Transgenic Plants,” Plant J. 11:605-612 (1997); and McNellis et al., “Glucocorticoid-Inducible Expression of a Bacterial Avirulence Gene in Transgenic Arabidopsis Induces Hypersensitive Cell Death, Plant J. 14(2):247-57 (1998), which are hereby incorporated by reference in their entirety). In addition, inducible promoters include promoters that function in a tissue specific manner to regulate the gene of interest within selected tissues of the plant. Examples of such tissue specific or developmentally regulated promoters include seed, flower, fruit, or root specific promoters as are well known in the field (U.S. Pat. No. 5,750,385 to Shewmaker et al., which is hereby incorporated by reference in its entirety).
  • A number of tissue- and organ-specific promoters have been developed for use in genetic engineering of plants (Potenza et al., “Targeting Transgene Expression in Research, Agricultural, and Environmental Applications: Promoters used in Plant Transformation,” In Vitro Cell. Dev. Biol. Plant 40:1-22 (2004), which is hereby incorporated by reference in its entirety). Examples of such promoters include those that are floral-specific (Annadana et al., “Cloning of the Chrysanthemum UEP1 Promoter and Comparative Expression in Florets and Leaves of Dendranthema grandiflora,” Transgenic Res. 11:437-445(2002), which is hereby incorporated by reference in its entirety), seed-specific (Kluth et al., “5′ Deletion of a gbss1 Promoter Region Leads to Changes in Tissue and Developmental Specificities,” Plant Mol. Biol. 49:669-682 (2002), which is hereby incorporated by reference in its entirety), root-specific (Yamamoto et al., “Characterization of cis-acting Sequences Regulating Root-Specific Gene Expression in Tobacco,” Plant Cell 3:371-382 (1991), which is hereby incorporated by reference in its entirety), fruit-specific (Fraser et al., “Evaluation of Transgenic Tomato Plants Expressing an Additional Phytoene Synthase in a Fruit-Specific Manner,” Proc. Natl Acad. Sci. USA 99:1092-1097 (2002), which is hereby incorporated by reference in its entirety), and tuber/storage organ-specific (Visser et al., “Expression of a Chimaeric Granule-Bound Starch Synthase-GUS gene in transgenic Potato Plants,” Plant Mol Biol. 17:691-699 (1991), which is hereby incorporated by reference in its entirety). Targeted expression of an introduced gene (transgene) is necessary when expression of the transgene could have detrimental effects if expressed throughout the plant. On the other hand, silencing a gene throughout a plant could also have negative effects. However, this problem could be avoided by localizing the silencing to a region by a tissue-specific promoter.
  • A suitable promoter can also be one that is gene-specific, in that it regulates transcription of a nucleic acid molecule of the present invention. A suitable gene-specific promoter gene-specific promoter (derived from MAGI93503) has a nucleotide sequence of SEQ ID NO:41 as follows:
  • CGTTGTCGGAACGTCCCGTCGATGTTCGGAAACGAGCACGACCCGTCGACTCCTGCTTCTTGGCGGAGA
    AGAAAGGGGACGACGAGCGAGCGTTTTGACTTTGATTTCCTCGCTAAAACCGGCCGCTGTTTTTGCTTT
    CCGCGCGAGCCGCCCACGTTATTGACTGACGCTGGTGCGAGAGCGCTGCTGCCTCTGCGGTTGCCGTCT
    GCGCTCCAGTGGTAGCCGAGAATATTGTTAGGTCCGTAGGATCAGATTTGCTACGTACTAAAAAAATTC
    CTTAAACTTTAATTGTGTATTTTTTTTAAAAAAAATTATAGCATTTATCAGCAACAAAACTCTAAAAAC
    ATGTTTAGTTCGCTGCTTAATTTATCACATATTGTCTAAATTTTATATATAAATTATTTAATTCGAACG
    ACTAACCAGAACCCAGACCTACAATAAATTTGCCCCCGCTGCTGCGCTCCCCAGCTCCCCAAGTCCCTA
    ACCCGCCCTCGCTTTGTCGCCGCGGCACACGGTTTTGGCCGTGGACAGGACAGTTGCACCCTAGCCCCA
    TTGGCCGATTCCGAGCTAGGAAGGAGTATATGCGTATCGGTAGTAACCGAGGAGCAACGCAACATGTCC
    ACAGCCCGCGCGCTGGTAACGGGTCCATGCGTCTTGGCTCATCAGGTGCCCCAAGGGACGCCCTCGCCC
    GGTCTGACCCACCTATATAAACTTAAAACTTGTGCCCCAACATCATCAGTTCGTATCACACCCAACCTC
    CCACTGTAAAAAAGAGCAGCGGAACGTGCGTGCATCCATCCATCCATCCATTTCCAATCCCAATCCCAA
    TCCCACCAGTGTCCAGTGCTCGGGGAACCGACACAGCTCCTCAGCAGAGTAGCCAGCACGACAAGCCCG
    ATCAGCAGACAGCAGGCATG

    This gene-specific promoter is a fragment of genomic DNA of maize that is likely to include promoter elements that allow the gene of SEQ ID NO:39 to exhibit nitrogen-regulated expression. Other suitable promoters include those having a nucleotide sequence of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and/or SEQ ID NO:46.
  • The nucleic acid construct of the present invention also includes an operable 3′ regulatory region, selected from among those which are capable of providing correct transcription termination and polyadenylation of mRNA for expression in the host cell of choice, operably linked to a modified trait nucleic acid molecule of the present invention. A number of 3′ regulatory regions are known to be operable in plants. Exemplary 3′ regulatory regions include, without limitation, the nopaline synthase (NOS) 3′ regulatory region (Fraley et al., “Expression of Bacterial Genes in Plant Cells,” Proc. Nat'l Acad. Sci. USA 80:4803-4807 (1983), which is hereby incorporated by reference in its entirety) and the cauliflower mosaic virus (CaMV) 3′ regulatory region (Odell et al., “Identification of DNA Sequences Required for Activity of the Cauliflower Mosaic Virus 35S Promoter,” Nature 313(6005):810-812 (1985), which is hereby incorporated by reference in its entirety). Virtually any 3′ regulatory region known to be operable in plants would be suitable for use in conjunction with the present invention.
  • The different components described above can be ligated together to produce expression systems which contain the nucleic acid constructs of the present invention, using well known molecular cloning techniques as described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition Cold Spring Harbor, N.Y.: Cold Spring Harbor Press (1989), and Ausubel et al. Current Protocols in Molecular Biology, New York, N.Y.: John Wiley & Sons (1989), which are hereby incorporated by reference in their entirety.
  • Once the nucleic acid construct of the present invention has been prepared, it is ready to be incorporated into a host cell. Accordingly, another aspect of the present invention relates to a recombinant host cell containing one or more of the nucleic acid constructs of the present invention. Basically, this method is carried out by transforming a host cell with a nucleic acid construct of the present invention under conditions effective to achieve transcription of the nucleic acid molecule in the host cell. This is achieved with standard cloning procedures known in the art, such as described by Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Springs Laboratory, Cold Springs Harbor, N.Y. (1989), which is hereby incorporated by reference in its entirety. Suitable hosts include, but are not limited to, bacterial cells, viruses, yeast cells, mammalian cells, insect cells, plant cells, and the like. Preferably the host is either a bacterial cell or a plant cell. Methods of transformation may result in transient or stable expression of the nucleic acid under control of the promoter. Preferably, a nucleic acid construct of the present invention is stably inserted into the genome of the recombinant plant cell as a result of the transformation, although transient expression can serve an important purpose, particularly when the plant under investigation is slow-growing.
  • Plant tissue suitable for transformation includes leaf tissue, root tissue, meristems, zygotic and somatic embryos, callus, protoplasts, tassels, pollen, embryos, anthers, and the like. The means of transformation chosen is that most suited to the tissue to be transformed.
  • Transient expression in plant tissue can be achieved by particle bombardment (Klein et al., “High-Velocity Microprojectiles for Delivering Nucleic Acids Into Living Cells,” Nature 327:70-73 (1987), which is hereby incorporated by reference in its entirety), also known as biolistic transformation of the host cell, as disclosed in U.S. Pat. Nos. 4,945,050, 5,036,006, and 5,100,792, all to Sanford et al., and in Emerschad et al., “Somatic Embryogenesis and Plant Development from Immature Zygotic Embryos of Seedless Grapes (Vitis vinifera),” Plant Cell Reports 14:6-12 (1995), which are hereby incorporated by reference in their entirety.
  • In particle bombardment, tungsten or gold microparticles (1 to 2 μm in diameter) are coated with the DNA of interest and then bombarded at the tissue using high pressure gas. In this way, it is possible to deliver foreign DNA into the nucleus and obtain a temporal expression of the gene under the current conditions of the tissue. Biologically active particles (e.g., dried bacterial cells containing the vector and heterologous DNA) can also be propelled into plant cells. Other variations of particle bombardment, now known or hereafter developed, can also be used. Further, particle bombardment transformation can be used to stably introduce the nucleic acid construct into plant cells.
  • Another appropriate method of stably introducing the nucleic acid construct into plant cells is to infect a plant cell with Agrobacterium tumefaciens or Agrobacterium rhizogenes previously transformed with the nucleic acid construct. As described above, the Ti (or RI) plasmid of Agrobacterium enables the highly successful transfer of a foreign nucleic acid molecule into plant cells. A variation of Agrobacterium transformation uses vacuum infiltration in which whole plants are used (Senior, “Uses of Plant Gene Silencing,” Biotechnology and Genetic Engineering Reviews 15:79-119 (1998), which is hereby incorporated by reference in its entirety).
  • Yet another method of introduction is fusion of protoplasts with other entities, either minicells, cells, lysosomes, or other fusible lipid-surfaced bodies (Fraley et al., Proc. Natl. Acad. Sci. USA 79:1859-63 (1982), which is hereby incorporated by reference in its entirety). The nucleic acid molecule may also be introduced into the plant cells by electroporation (Fromm et al., Proc. Natl Acad. Sci. USA 82:5824 (1985), which is hereby incorporated by reference in its entirety). In this technique, plant protoplasts are electroporated in the presence of plasmids containing the expression cassette. Electrical impulses of high field strength reversibly permeabilize biomembranes allowing the introduction of the plasmids. Electroporated plant protoplasts reform the cell wall, divide, and regenerate. Other methods of transformation include polyethylene-mediated plant transformation, micro-injection, physical abrasives, and laser beams (Senior, “Uses of Plant Gene Silencing,” Biotechnology and Genetic Engineering Reviews 15:79-119 (1998), which is hereby incorporated by reference in its entirety). The precise method of transformation is not critical to the practice of the present invention. Any method that results in efficient transformation of the host cell of choice is appropriate for practicing the present invention. Transfromation can also be achieved using the “whisker” method, as is well known in the art.
  • After transformation, the transformed plant cells must be regenerated. Plant regeneration from cultured protoplasts is described in Evans et al., Handbook of Plant Cell Cultures, Vol 1, New York, N.Y.: MacMillan Publishing Co. (1983); Vasil, ed., Cell Culture and Somatic Cell Genetics of Plants, Vol. I (1984) and Vol. III (1986), Orlando: Acad. Press, which are hereby incorporated by reference in their entirety.
  • Means for regeneration vary from species to species of plants, but generally a suspension of transformed protoplasts or a petri plate containing explants is first provided. Callus tissue is formed and shoots may be induced from callus and subsequently rooted. Alternatively, embryo formation can be induced in the callus tissue. These embryos germinate as natural embryos to form plants. The culture media will generally contain various amino acids and hormones, such as auxin and cytokinins. Efficient regeneration will depend on the medium, on the genotype, and on the history of the culture. If these three variables are controlled, then regeneration is usually reproducible and repeatable.
  • Preferably, transformed cells are first identified using a selection marker simultaneously introduced into the host cells along with the nucleic acid construct of the present invention. Suitable selection markers include, without limitation, markers encoding for antibiotic resistance, such as the neomycin phosphotransferae II (“nptII”) gene which confers kanamycin resistance (Fraley et al., Proc. Natl. Acad. Sci. USA 80:4803-4807 (1983), which is hereby incorporated by reference in its entirety), and the genes which confer resistance to gentamycin, G418, hygromycin, streptomycin, spectinomycin, tetracycline, chloramphenicol, and the like. Cells or tissues are grown on a selection medium containing the appropriate antibiotic, whereby generally only those transformants expressing the antibiotic resistance marker continue to grow. Other types of markers are also suitable for inclusion in the expression cassette of the present invention. For example, a gene encoding for herbicide tolerance, such as tolerance to sulfonylurea is useful, or the dhfr gene, which confers resistance to methotrexate (Bourouis et al., EMBO J. 2:1099-1104 (1983), which is hereby incorporated by reference in its entirety). Similarly, “reporter genes,” which encode for enzymes providing for production of an identifiable compound are suitable. The most widely used reporter gene for gene fusion experiments has been uidA, a gene from Escherichia coli that encodes the β-glucuronidase protein, also known as GUS (Jefferson et al., “GUS Fusions: β Glucuronidase as a Sensitive and Versatile Gene Fusion Marker in Higher Plants,” EMBO J 6:3901-3907 (1987), which is hereby incorporated by reference in its entirety). Similarly, enzymes providing for production of a compound identifiable by luminescence, such as luciferase, are useful. The selection marker employed will depend on the target species; for certain target species, different antibiotics, herbicide, or biosynthesis selection markers are preferred.
  • Plant cells and tissues selected by means of an inhibitory agent or other selection marker are then tested for the acquisition of the transgene (Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, N.Y.: Cold Spring Harbor Press (1989), which is hereby incorporated by reference in its entirety).
  • After the fusion gene containing a nucleic acid construct of the present invention is stably incorporated in transgenic plants, the transgene can be transferred to other plants by sexual crossing. Any of a number of standard breeding techniques 10 can be used, depending upon the species to be crossed. Once transgenic plants of this type are produced, the plants themselves can be cultivated in accordance with conventional procedure so that the nucleic acid construct is present in the resulting plants. Alternatively, transgenic seeds are recovered from the transgenic plants. These seeds can then be planted in the soil and cultivated using conventional procedures to produce transgenic plants. The component parts and fruit of such plants are encompassed by the present invention.
  • The present invention can be utilized in conjunction with a wide variety of plants or their seeds. Suitable plants can include dicots and monocots. More particular, suitable plants can include the following: rice, corn, soybean, canola, potato, wheat, mung bean, alfalfa, barley, rye, cotton, sunflower, peanut, sweet potato, bean, pea, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, tobacco, tomato, sorghum, sugarcane, banana, Arabidopsis thaliana, Saintpaulia, petunia, pelargonium, poinsettia, chrysanthemum, carnation, crocus, marigold, daffodil, pine, Medicago truncatula, Sandersonlia aurantiaca, and zinnia.
  • Another aspect of the present invention is a method of expressing a nucleic acid molecule that is modulated by nitrogen in a plant. This method involves providing a transgenic plant or plant seed transformed with a nucleic acid construct having a nucleic acid molecule that is modulated by nitrogen in corn, a 5′ DNA promoter sequence, and a 3′ terminator sequence. The nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule. The method also involves growing the transgenic plant or a transgenic plant grown from the transgenic plant seed under conditions effective to express the nucleic acid molecule in the transgenic plant or the plant grown from the transgenic plant seed. In one embodiment, the transgenic plant or plant seed is provided by transforming a non-transgenic plant or a non-transgenic plant seed with the nucleic acid construct of the present invention to yield said transgenic plant or plant seed. In one aspect, the growing step is effective in reducing nitrogen uptake of the transgenic plant or the plant grown from the transgenic plant seed. In another aspect, the growing step is effective in increasing nitrogen uptake of the transgenic plant or the plant grown from the transgenic plant seed. In yet another aspect, the growing step is effective in increasing efficiency of nitrogen utilization of the transgenic plant or the plant grown from the transgenic plant seed. Transformation of the transgenic plant or plant seed can be achieved using Agrobacterium-mediated transformation, the whisker method, vacuum infiltration, biolistic transformation, electroporation, micro-injection, polyethylene-mediated transformation, or laser-beam transformation.
  • The present invention also relates to an isolated DNA promoter from corn suitable for inducing nitrogen-regulated expression of a protein encoded by an isolated DNA molecule operably associated with the DNA promoter. A suitable DNA promoter for use in this method can be any one of the promoters described herein, including, for example, the promoters having a nucleotide sequence of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and/or SEQ ID NO:46. The isolated DNA promoter can be used to prepare nucleic acid constructs as previously described. In a particular nucleic acid construct, the isolated DNA promoter can be operably linked to an isolated nucleic acid that either has a nucleotide sequence (or encoding portion thereof) of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and/or SEQ ID NO:44, or encodes a polypeptide having an amino acid sequence of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:28, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:40, SEQ ID NO:43, and/or SEQ ID NO:45. Other suitable genes from corn that can be regulated by the DNA promoter of the present invention include, for example, nitrate reductase, nitrite reductase, Uroporphyrin-III methyl transferase. Expression vectors can be prepared by inserting the nucleic acid construct in an appropriate vector (as described in more detail supra), and transgenic host cells and plants (including their component parts such as fruits and seeds) can be produced by transforming them with the nucleic acid construct containing the DNA promoter.
  • The present invention also relates to a method of directing nitrogen-regulated expression of an isolated nucleic acid in plants. This methods involves transforming a plant cell with the nucleic acid construct that includes an isolated DNA promoter suitable for inducing nitrogenregulated expression of a protein encoded by an isolated DNA molecule operably associated with the DNA promoter. This method also involves regenerating a plant from the transformed plant cell. By this method, expression of the nucleic acid molecule, under control of the DNA promoter, occurs in the plant and is upregulated by nitrogen. A suitable DNA promoter for use in this method can be any one of the promoters described herein, including, for example, the promoters having a nucleotide sequence of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and/or SEQ ID NO:46.
  • Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be made without departing from the spirit of the invention and these are, therefore, considered to be within the scope of the invention as defined in the claims which follow.

Claims (37)

1. A nucleic acid construct comprising:
a nucleic acid molecule that is modulated by nitrogen in corn;
a 5′ DNA promoter sequence; and
a 3′ terminator sequence, wherein the nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule.
2. The nucleic acid construct according to claim 1, wherein the nucleic acid molecule either:
(a) encodes a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:28, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:40, SEQ ID NO:43, and SEQ ID NO:45; or
(b) comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and SEQ ID NO:44; or
(c) comprises a coding portion of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and SEQ ID NO:44.
3. The nucleic acid construct according to claim 1, wherein the DNA promoter sequence is a constitutive plant promoter.
4. The nucleic acid construct according to claim 1, wherein the DNA promoter sequence is an inducible plant promoter.
5. The nucleic acid construct according to claim 4, wherein the inducible plant promoter is a nitrogen inducible plant promoter.
6. The nucleic acid construct according to claim 5, wherein the nitrogen inducible plant promoter comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and SEQ ID NO:46.
7. The nucleic acid construct according to claim 1, wherein the DNA promoter sequence is a tissue-specific promoter.
8. The nucleic acid construct according to claim 1, wherein the DNA promoter sequence is an organ-specific promoter.
9. An expression vector comprising the nucleic acid construct according to claim 1.
10. A host cell transformed with the nucleic acid construct according to claim 1.
11. The host cell according to claim 10, wherein the host cell is a bacterial cell or a plant cell.
12. The host cell according to claim 11, wherein the host cell is a plant cell.
13. A plant transformed with the nucleic acid construct according to claim 1.
14. The plant according to claim 13, wherein the plant is selected from the group consisting of nice, corn, soybean, canola, potato, wheat, mung bean, alfalfa, barley, rye, cotton, sunflower, peanut, sweet potato, bean, pea, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, tobacco, tomato, sorghum, sugarcane, banana, Arabidopsis thaliana, Saintpaulia, petunia, pelargonium, poinsettia, chrysanthemum, carnation, crocus, marigold, daffodil, pine, Medicago truncatula, Sandersonia aurantiaca, and zinnia.
15. A component part of the plant according to claim 13.
16. A fruit of the plant according to claim 13.
17. A plant seed produced from the plant according to claim 13.
18. A plant seed transformed with the nucleic acid construct according to claim 1.
19. A method of expressing a nucleic acid molecule modulated by nitrogen in a plant, said method comprising:
providing a transgenic plant or plant seed transformed with a nucleic acid construct according to claim 1 and growing the transgenic plant or a plant grown from the transgenic plant seed under conditions effective to express the nucleic acid molecule in said transgenic plant or said plant grown from the transgenic plant seed.
20. The method according to claim 19, wherein said growing is effective in reducing nitrogen uptake of said transgenic plant or said plant grown from the transgenic plant seed.
21. The method according to claim 19, wherein said growing is effective in increasing nitrogen uptake of said transgenic plant or said plant grown from the transgenic plant seed.
22. The method according to claim 19, wherein said growing is effective in increasing efficiency of nitrogen utilization of said transgenic plant or said plant grown from the transgenic plant seed.
23. The method according to claim 19, wherein a transgenic plant is provided.
24. The method according to claim 19, wherein a transgenic plant seed is provided.
25. The method according to claim 19, wherein the plant is selected from the group consisting of rice, corn, soybean, canola, potato, wheat, mung bean, alfalfa, barley, rye, cotton, sunflower, peanut, sweet potato, bean, pea, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, tobacco, tomato, sorghum, sugarcane, banana, Arabidopsis thaliana, Saintpaulia, petunia, pelargonium, poinsettia, chrysanthemum, carnation, crocus, marigold, daffodil, pine, Medicago truncatula, Sandersonia aurantiaca, and zinnia.
26. The method according to claim 19, wherein said providing comprises transforming a non-transgenic plant or a non-transgenic plant seed with the nucleic acid construct to yield said transgenic plant or plant seed.
27. The method according to claim 26, wherein said transforming comprises Agrobacterium-mediated transformation, whisker method transformation, vacuum infiltration, biolistic transformation, electroporation, micro-injection, polyethylene-mediated transformation, or laser-beam transformation.
28. An isolated DNA promoter from corn suitable for inducing nitrogen-regulated expression of a protein encoded by an isolated DNA molecule operably associated with the DNA promoter, wherein said DNA promoter comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and SEQ ID NO:46.
29. A nucleic acid construct comprising:
an isolated nucleic acid molecule encoding a protein;
an isolated DNA promoter according to claim 28, wherein the DNA promoter is operably linked 5′ to the isolated nucleic acid molecule to induce transcription of the nucleic acid molecule; and
a 3′ terminator sequence operably linked to the isolated nucleic acid molecule.
30. An expression vector comprising the nucleic acid construct according to claim 29.
31. A host cell transformed with the nucleic acid construct according to claim 29.
32. A plant transformed with the nucleic acid construct according to claim 29.
33. A plant seed produced from the plant according to claim 32.
34. A plant seed transformed with the nucleic acid construct according to claim 29.
35. A method of directing nitrogen-regulated expression of an isolated nucleic acid in plants, said method comprising:
transforming a plant cell with a nucleic acid construct according to claim 29 and
regenerating a plant from the transformed plant cell, wherein expression of the nucleic acid molecule, under control of the DNA promoter, occurs in the plant and is upregulated by nitrogen.
36. An isolated nucleic acid molecule that is modulated by nitrogen in corn, wherein said nucleic acid molecule comprises:
a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and SEQ ID NO:44; or
a coding portion of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and SEQ ID NO:44.
37. An isolated protein or polypeptide, wherein said isolated protein or polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:28, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:40, SEQ ID NO:43, and SEQ ID NO:45.
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130313918A1 (en) * 2011-02-02 2013-11-28 Siemens Aktiengesellschaft Method for protecting charging cable, and charging device
CN104711260A (en) * 2013-12-13 2015-06-17 华中农业大学 Promoter Y8A for specific induced expression by recovering nitrogen supply after nitrogen deficiency of rice and application thereof
CN108410888A (en) * 2018-03-15 2018-08-17 山东农业大学 A kind of apple MdCEPR1 genes and its preparation method and application

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013152317A2 (en) * 2012-04-06 2013-10-10 Syngenta Participations Ag Nitrite transporter and methods of using the same
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Citations (91)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5391725A (en) * 1989-12-08 1995-02-21 New York University Medical Center Organ-specific plant promoter sequences
US5608144A (en) * 1994-08-12 1997-03-04 Dna Plant Technology Corp. Plant group 2 promoters and uses thereof
US5633441A (en) * 1989-08-04 1997-05-27 Plant Genetic Systems, N.V. Plants with genetic female sterility
US6084153A (en) * 1996-02-14 2000-07-04 The Governors Of The University Of Alberta Plants having enhanced nitrogen assimilation/metabolism
US6107547A (en) * 1993-10-06 2000-08-22 New York University Transgenic plants that exhibit enhanced nitrogen assimilation
US20010000266A1 (en) * 1995-10-06 2001-04-12 Schmidt Robert R. Novel polypeptides and polynucleotides relating to the alpha- and beta-subunits of glutamate dehydrogenases and methods of use
US20010003848A1 (en) * 1993-10-28 2001-06-14 Wolf-Bernd Frommer Dna sequences for ammonium transporter, plasmids, bacteria, yeasts, plant cells and plants containing the transporter
US6342581B1 (en) * 1997-07-08 2002-01-29 Human Genome Sciences, Inc. Secreted protein HLHFP03
US20020069430A1 (en) * 1993-10-06 2002-06-06 Ajinomoto Co., Inc. Transgenic plants that exhibit enhanced nitrogen assimilation
US6506963B1 (en) * 1999-12-08 2003-01-14 Plant Genetic Systems, N.V. Hybrid winter oilseed rape and methods for producing same
US20030022305A1 (en) * 1995-06-07 2003-01-30 New York University. Plant glutamate receptors
US20040031072A1 (en) * 1999-05-06 2004-02-12 La Rosa Thomas J. Soy nucleic acid molecules and other molecules associated with transcription plants and uses thereof for plant improvement
US20040034888A1 (en) * 1999-05-06 2004-02-19 Jingdong Liu Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US20040123343A1 (en) * 2000-04-19 2004-06-24 La Rosa Thomas J. Rice nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US6852519B2 (en) * 2000-06-06 2005-02-08 Applera Corporation Isolated human kinase proteins, nucleic acid molecules encoding human kinase proteins, and uses thereof
US20050158323A1 (en) * 2003-12-04 2005-07-21 Vaccinex, Inc. Methods of killing tumor cells by targeting internal antigens exposed on apoptotic tumor cells
US6936393B2 (en) * 2001-12-06 2005-08-30 Kao Corporation Catalyst for preparing polyester for toner
US6939701B2 (en) * 2003-08-08 2005-09-06 Novozymes, Inc. Polypeptides having oxaloacetate hydrolase activity and nucleic acids encoding same
US6942973B2 (en) * 2001-03-12 2005-09-13 Novozymes Biotech, Inc. Methods for isolating genes from microorganisms
US20060048240A1 (en) * 2004-04-01 2006-03-02 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US7038030B2 (en) * 2002-04-16 2006-05-02 University Of South Florida BIVM (basic, immunoglobulin-like variable motif-containing) gene, transcriptional products, and uses thereof
US20060107345A1 (en) * 2003-09-30 2006-05-18 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20060123505A1 (en) * 2002-05-30 2006-06-08 National Institute Of Agrobiological Sciences Full-length plant cDNA and uses thereof
US20060150283A1 (en) * 2004-02-13 2006-07-06 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20070016974A1 (en) * 1999-09-30 2007-01-18 Byrum Joseph R Nucleic acid molecules and other molecules associated with plants
US20070020621A1 (en) * 2000-07-19 2007-01-25 Boukharov Andrey A Genomic plant sequences and uses thereof
US20070061916A1 (en) * 2001-05-07 2007-03-15 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US7193033B2 (en) * 2000-06-21 2007-03-20 Takeda Pharmaceutical Company Limited Peptide having appetite stimulating activity
US20070067865A1 (en) * 2000-09-05 2007-03-22 Kovalic David K Annotated plant genes
US7195901B1 (en) * 2004-06-03 2007-03-27 The United States Of America As Represented By The Secretary Of Agriculture Diacylglycerol acyltransferase and its use to preferentially incorporate fatty acids into diacylglycerol
US7214786B2 (en) * 2000-12-14 2007-05-08 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US20070143875A1 (en) * 2000-04-03 2007-06-21 Conner Timothy W Polynucleotides and Other Molecules Associated With Plants
US20070150978A1 (en) * 1999-05-17 2007-06-28 Byrum Joseph R Nucleic acid molecules and other molecules associated with plants
US7247440B2 (en) * 2001-05-15 2007-07-24 Takeda Pharmaceutical Company Limited Method of screening preventives or remedies for obesity
US7348142B2 (en) * 2002-03-29 2008-03-25 Veridex, Lcc Cancer diagnostic panel
US20080114160A1 (en) * 2000-10-31 2008-05-15 Andrey Boukharov Plant Genome Sequence and Uses Thereof
US7375074B2 (en) * 2001-12-28 2008-05-20 Takeda Chemical Industries, Ltd. Body weight gain inhibitor
US7396979B2 (en) * 2004-06-30 2008-07-08 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying plant characteristics and phenotypes
US7399587B2 (en) * 2003-05-29 2008-07-15 Astellas Pharma Inc. Canine CYP1A2 genetic polymorphism
US20080206837A1 (en) * 2002-04-25 2008-08-28 Crucell Holland B.V. Stable adenoviral vectors and methods for propagation thereof
US7473526B2 (en) * 2002-03-29 2009-01-06 Veridex, Llc Breast cancer prognostic portfolio
US7507875B2 (en) * 2003-06-06 2009-03-24 Arborgen, Llc Transcription factors
US7511190B2 (en) * 1999-11-17 2009-03-31 Mendel Biotechnology, Inc. Polynucleotides and polypeptides in plants
US20090087878A9 (en) * 1999-05-06 2009-04-02 La Rosa Thomas J Nucleic acid molecules associated with plants
US7514597B2 (en) * 2002-08-20 2009-04-07 Suntory Limited Glycosyltransferase gene
US20090094717A1 (en) * 2007-10-03 2009-04-09 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics
US20090093620A1 (en) * 2000-09-05 2009-04-09 David Kovalic Annotated Plant Genes
US7521230B2 (en) * 2002-10-16 2009-04-21 Board Of Regents Of The University Of Nebraska Nucleic acid encoding a Brain Derived Tau Kinase polypeptide and methods of use thereof
US7525016B1 (en) * 2004-05-21 2009-04-28 Iowa State University Research Foundation, Inc. Identification of syn-copalyl diphosphate synthase
US20090123986A1 (en) * 2002-06-21 2009-05-14 Divergence, Inc. Nematode Phosphoethanolamine N-Methyltransferase-Like Sequences
US7538204B2 (en) * 2004-01-21 2009-05-26 Malaysian Palm Oil Board Recombinant enzyme and uses therefor
US20090144848A1 (en) * 2000-05-08 2009-06-04 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US7569389B2 (en) * 2004-09-30 2009-08-04 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying plant characteristics
US20090208564A1 (en) * 2007-08-27 2009-08-20 Chiang Jia Li Compositions of asymmetric interfering RNA and uses thereof
US20100011463A1 (en) * 2007-02-06 2010-01-14 Basf Plant Science Gmbh Compositions and Methods Using RNA Interference for Control of Nematodes
US20100017910A1 (en) * 2007-02-09 2010-01-21 Basf Plant Science Gmbh Compositions and Methods Using RNA Interference of CDPK-Like For Control of Nematodes
US7659446B2 (en) * 2003-02-25 2010-02-09 Mendel Biotechnology, Inc. Polynucleotides and polypeptides in plants
US7700852B2 (en) * 2002-05-06 2010-04-20 Jeroen Demmer Compositions isolated from forage grasses and methods for their use
US7709611B2 (en) * 2004-08-04 2010-05-04 Amgen Inc. Antibodies to Dkk-1
US20100162433A1 (en) * 2006-10-27 2010-06-24 Mclaren James Plants with improved nitrogen utilization and stress tolerance
US7745391B2 (en) * 2001-09-14 2010-06-29 Compugen Ltd. Human thrombospondin polypeptide
US20100168205A1 (en) * 2008-10-23 2010-07-01 Alnylam Pharmaceuticals, Inc. Methods and Compositions for Prevention or Treatment of RSV Infection Using Modified Duplex RNA Molecules
US7750207B2 (en) * 2004-09-01 2010-07-06 Monsanto Technology Llc Zea mays ribulose bisphosphate carboxylase activase promoter
US7750209B2 (en) * 2003-12-17 2010-07-06 International Flower Developments Proprietary Limited Method for producing yellow flower by controlling flavonoid synthetic pathway
US7786272B2 (en) * 1998-03-27 2010-08-31 Genentech, Inc. Antibodies to PRO352
US7863001B2 (en) * 2001-06-18 2011-01-04 The Netherlands Cancer Institute Diagnosis and prognosis of breast cancer patients
US7867749B2 (en) * 2002-06-21 2011-01-11 Divergence, Inc. Nematode phosphoethanolamine N-methyltransferase-like sequences
US7868149B2 (en) * 1999-07-20 2011-01-11 Monsanto Technology Llc Plant genome sequence and uses thereof
US20110008322A1 (en) * 2007-12-26 2011-01-13 Vaccinex, Inc. Anti-c35 antibody combination therapies and methods
US7892730B2 (en) * 2000-12-22 2011-02-22 Sagres Discovery, Inc. Compositions and methods for cancer
US7910800B2 (en) * 2005-08-15 2011-03-22 Evogene Ltd. Methods of increasing abiotic stress tolerance and/or biomass in plants and plants generated thereby
US20110105347A1 (en) * 2007-05-17 2011-05-05 Monsanto Technology Llc Corn polymorphisms and methods of genotyping
US20110126316A1 (en) * 2008-05-01 2011-05-26 Academia Sinica Use of rice polypeptides/nucleic acids for plant improvement
US7956242B2 (en) * 1998-09-22 2011-06-07 Mendel Biotechnology, Inc. Plant quality traits
US20110138499A1 (en) * 1999-03-23 2011-06-09 Mendel Biotechnology, Inc. Plant quality traits
US7960612B2 (en) * 1998-09-22 2011-06-14 Mendel Biotechnology, Inc. Plant quality with various promoters
US20110145946A1 (en) * 2008-08-18 2011-06-16 Evogene Ltd. Isolated polypeptides and polynucleotides useful for increasing nitrogen use efficiency, abiotic stress tolerance, yield and biomass in plants
US7968689B2 (en) * 1997-03-07 2011-06-28 Human Genome Sciences, Inc. Antibodies to HSDEK49 polypeptides
US20110158904A1 (en) * 2002-10-16 2011-06-30 Corixa Corporation Antibodies that bind cell-associated ca 125/o772p and methods of use thereof
US20110167514A1 (en) * 2007-07-05 2011-07-07 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics
US20110177228A1 (en) * 2008-08-13 2011-07-21 Ceres, Inc. Plant nucelotide sequences and corresponding polypeptides
US20110179501A1 (en) * 2007-07-31 2011-07-21 The Ohio State University Research Foundation Methods for Reverting Methylation by Targeting DNMT3A and DNMT3B
US20110179531A1 (en) * 2000-05-09 2011-07-21 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US7985571B1 (en) * 2002-06-07 2011-07-26 Ryogen Llc Isolated genomic polynucleotide fragments from chromosome 17 that encode human carboxypeptidase D
US20110184045A1 (en) * 2008-06-30 2011-07-28 Gunther Hartmann Silencng and rig-i activation by dual function oligonucleotides
US7989676B2 (en) * 2006-08-31 2011-08-02 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics
US20110201667A1 (en) * 2009-07-20 2011-08-18 Protiva Biotherapeutics, Inc. Compositions and methods for silencing ebola virus gene expression
US20120017338A1 (en) * 2008-01-15 2012-01-19 Wei Wu Isolated novel nucleic acid and protein molecules from corn and methods of using those molecules to generate transgenic plant with enhanced agronomic traits
US20120017292A1 (en) * 2009-01-16 2012-01-19 Kovalic David K Isolated novel nucleic acid and protein molecules from corn and methods of using those molecules to generate transgene plants with enhanced agronomic traits
US20120023611A1 (en) * 2009-02-27 2012-01-26 Yongwei Cao Isolated Novel Nucleic Acid and Protein Molecules from Corn and Methods of Using Thereof
US20120159672A1 (en) * 2004-02-06 2012-06-21 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby

Family Cites Families (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7390937B2 (en) 1996-02-14 2008-06-24 The Governors Of The University Of Alberta Plants with enhanced levels of nitrogen utilization proteins in their root epidermis and uses thereof
US5998700A (en) 1996-07-02 1999-12-07 The Board Of Trustees Of Southern Illinois University Plants containing a bacterial Gdha gene and methods of use thereof
US8030546B2 (en) 1998-09-22 2011-10-04 Mendel Biotechnology, Inc. Biotic and abiotic stress tolerance in plants
US20110214206A1 (en) 1999-05-06 2011-09-01 La Rosa Thomas J Nucleic acid molecules and other molecules associated with plants
US7834146B2 (en) 2000-05-08 2010-11-16 Monsanto Technology Llc Recombinant polypeptides associated with plants
US20070224595A1 (en) 2000-06-15 2007-09-27 Andersen Scott E Nucleic acid molecules and other molecules associated with plants
USH2220H1 (en) 2001-08-10 2008-07-01 Snp Consortium Identification and mapping of single nucleotide polymorphisms in the human genome
AU2002361610B2 (en) 2001-11-07 2007-01-11 Agensys, Inc. Nucleic acid and corresponding protein entitled 161P2F10B useful in treatment and detection of cancer
US7439067B2 (en) 2002-05-20 2008-10-21 Battelle Memorial Institute Isolated polynucleotides and methods of promoting a morphology in a fungus
US6936693B2 (en) 2003-02-04 2005-08-30 Divergence, Inc. Nematode PAN and ZP receptor-like sequences
EP2365063B1 (en) 2003-03-31 2015-10-14 University Of Bristol New vegetable acyltransferases specifically for long-chain polyunsaturated fatty acids
US20070277269A1 (en) 2006-04-17 2007-11-29 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying plant characteristics
IL164125A (en) 2004-09-19 2014-06-30 Arthur Schaffer Gene controlling fissure development in tomato fruit cuticles
US7838278B2 (en) 2005-01-14 2010-11-23 Archer-Daniels-Midland Company Compositions and methods for manipulating carbon flux in cells
US7435168B2 (en) 2005-01-14 2008-10-14 Archer-Daniels-Midland Company Compositions and methods for manipulating carbon flux in cells
US8014957B2 (en) 2005-12-15 2011-09-06 Fred Hutchinson Cancer Research Center Genes associated with progression and response in chronic myeloid leukemia and uses thereof
CA2681661C (en) 2007-03-23 2015-11-24 New York University Methods of increasing nitrogen-assimilation capacity in transgenic plants expressing cca1 and glk1
US7611705B2 (en) 2007-06-15 2009-11-03 National Cheng Kung University Anti-IL-20 antibody and its use in treating IL-20 associated inflammatory diseases
US8030290B2 (en) 2007-12-07 2011-10-04 City Of Hope Cell-type specific aptamer-siRNA delivery system for HIV-1 Therapy
AU2008334948B2 (en) 2007-12-13 2014-11-20 Alnylam Pharmaceuticals, Inc. Methods and compositions for prevention or treatment of RSV infection
US20110214205A1 (en) 2010-02-26 2011-09-01 Monsanto Technology Llc. Isolated Novel Nucleic Acid and Protein Molecules from Foxtail Millet and Methods of Using Those Molecules to Generate Transgenic Plants with Enhanced Agronomic Traits

Patent Citations (99)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5633441A (en) * 1989-08-04 1997-05-27 Plant Genetic Systems, N.V. Plants with genetic female sterility
US5391725A (en) * 1989-12-08 1995-02-21 New York University Medical Center Organ-specific plant promoter sequences
US20020069430A1 (en) * 1993-10-06 2002-06-06 Ajinomoto Co., Inc. Transgenic plants that exhibit enhanced nitrogen assimilation
US6107547A (en) * 1993-10-06 2000-08-22 New York University Transgenic plants that exhibit enhanced nitrogen assimilation
US20010003848A1 (en) * 1993-10-28 2001-06-14 Wolf-Bernd Frommer Dna sequences for ammonium transporter, plasmids, bacteria, yeasts, plant cells and plants containing the transporter
US5608144A (en) * 1994-08-12 1997-03-04 Dna Plant Technology Corp. Plant group 2 promoters and uses thereof
US20030022305A1 (en) * 1995-06-07 2003-01-30 New York University. Plant glutamate receptors
US20010000266A1 (en) * 1995-10-06 2001-04-12 Schmidt Robert R. Novel polypeptides and polynucleotides relating to the alpha- and beta-subunits of glutamate dehydrogenases and methods of use
US6084153A (en) * 1996-02-14 2000-07-04 The Governors Of The University Of Alberta Plants having enhanced nitrogen assimilation/metabolism
US7968689B2 (en) * 1997-03-07 2011-06-28 Human Genome Sciences, Inc. Antibodies to HSDEK49 polypeptides
US6342581B1 (en) * 1997-07-08 2002-01-29 Human Genome Sciences, Inc. Secreted protein HLHFP03
US7786272B2 (en) * 1998-03-27 2010-08-31 Genentech, Inc. Antibodies to PRO352
US7956242B2 (en) * 1998-09-22 2011-06-07 Mendel Biotechnology, Inc. Plant quality traits
US7960612B2 (en) * 1998-09-22 2011-06-14 Mendel Biotechnology, Inc. Plant quality with various promoters
US20110138499A1 (en) * 1999-03-23 2011-06-09 Mendel Biotechnology, Inc. Plant quality traits
US20040031072A1 (en) * 1999-05-06 2004-02-12 La Rosa Thomas J. Soy nucleic acid molecules and other molecules associated with transcription plants and uses thereof for plant improvement
US20040034888A1 (en) * 1999-05-06 2004-02-19 Jingdong Liu Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US20090087878A9 (en) * 1999-05-06 2009-04-02 La Rosa Thomas J Nucleic acid molecules associated with plants
US20070150978A1 (en) * 1999-05-17 2007-06-28 Byrum Joseph R Nucleic acid molecules and other molecules associated with plants
US7868149B2 (en) * 1999-07-20 2011-01-11 Monsanto Technology Llc Plant genome sequence and uses thereof
US20070016974A1 (en) * 1999-09-30 2007-01-18 Byrum Joseph R Nucleic acid molecules and other molecules associated with plants
US7511190B2 (en) * 1999-11-17 2009-03-31 Mendel Biotechnology, Inc. Polynucleotides and polypeptides in plants
US8110725B2 (en) * 1999-11-17 2012-02-07 Mendel Biotechnology, Inc. Polynucleotides and polypeptides in plants
US6506963B1 (en) * 1999-12-08 2003-01-14 Plant Genetic Systems, N.V. Hybrid winter oilseed rape and methods for producing same
US20070143875A1 (en) * 2000-04-03 2007-06-21 Conner Timothy W Polynucleotides and Other Molecules Associated With Plants
US20040123343A1 (en) * 2000-04-19 2004-06-24 La Rosa Thomas J. Rice nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US8106174B2 (en) * 2000-05-08 2012-01-31 Monsanto Technology Llc Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US20090144848A1 (en) * 2000-05-08 2009-06-04 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US20110179531A1 (en) * 2000-05-09 2011-07-21 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US6852519B2 (en) * 2000-06-06 2005-02-08 Applera Corporation Isolated human kinase proteins, nucleic acid molecules encoding human kinase proteins, and uses thereof
US7193033B2 (en) * 2000-06-21 2007-03-20 Takeda Pharmaceutical Company Limited Peptide having appetite stimulating activity
US20070020621A1 (en) * 2000-07-19 2007-01-25 Boukharov Andrey A Genomic plant sequences and uses thereof
US20090093620A1 (en) * 2000-09-05 2009-04-09 David Kovalic Annotated Plant Genes
US20070067865A1 (en) * 2000-09-05 2007-03-22 Kovalic David K Annotated plant genes
US20080114160A1 (en) * 2000-10-31 2008-05-15 Andrey Boukharov Plant Genome Sequence and Uses Thereof
US20110209246A1 (en) * 2000-12-14 2011-08-25 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US7214786B2 (en) * 2000-12-14 2007-05-08 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US7892730B2 (en) * 2000-12-22 2011-02-22 Sagres Discovery, Inc. Compositions and methods for cancer
US6942973B2 (en) * 2001-03-12 2005-09-13 Novozymes Biotech, Inc. Methods for isolating genes from microorganisms
US20070061916A1 (en) * 2001-05-07 2007-03-15 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US7247440B2 (en) * 2001-05-15 2007-07-24 Takeda Pharmaceutical Company Limited Method of screening preventives or remedies for obesity
US7863001B2 (en) * 2001-06-18 2011-01-04 The Netherlands Cancer Institute Diagnosis and prognosis of breast cancer patients
US7745391B2 (en) * 2001-09-14 2010-06-29 Compugen Ltd. Human thrombospondin polypeptide
US6936393B2 (en) * 2001-12-06 2005-08-30 Kao Corporation Catalyst for preparing polyester for toner
US7375074B2 (en) * 2001-12-28 2008-05-20 Takeda Chemical Industries, Ltd. Body weight gain inhibitor
US7348142B2 (en) * 2002-03-29 2008-03-25 Veridex, Lcc Cancer diagnostic panel
US7473526B2 (en) * 2002-03-29 2009-01-06 Veridex, Llc Breast cancer prognostic portfolio
US7038030B2 (en) * 2002-04-16 2006-05-02 University Of South Florida BIVM (basic, immunoglobulin-like variable motif-containing) gene, transcriptional products, and uses thereof
US20080206837A1 (en) * 2002-04-25 2008-08-28 Crucell Holland B.V. Stable adenoviral vectors and methods for propagation thereof
US7700852B2 (en) * 2002-05-06 2010-04-20 Jeroen Demmer Compositions isolated from forage grasses and methods for their use
US20060123505A1 (en) * 2002-05-30 2006-06-08 National Institute Of Agrobiological Sciences Full-length plant cDNA and uses thereof
US7985571B1 (en) * 2002-06-07 2011-07-26 Ryogen Llc Isolated genomic polynucleotide fragments from chromosome 17 that encode human carboxypeptidase D
US7867749B2 (en) * 2002-06-21 2011-01-11 Divergence, Inc. Nematode phosphoethanolamine N-methyltransferase-like sequences
US20090123986A1 (en) * 2002-06-21 2009-05-14 Divergence, Inc. Nematode Phosphoethanolamine N-Methyltransferase-Like Sequences
US7514597B2 (en) * 2002-08-20 2009-04-07 Suntory Limited Glycosyltransferase gene
US20110158904A1 (en) * 2002-10-16 2011-06-30 Corixa Corporation Antibodies that bind cell-associated ca 125/o772p and methods of use thereof
US20100058489A1 (en) * 2002-10-16 2010-03-04 Tsuneya Ikezu Nucleic Acid Encoding A Brain Derived Tau Kinase Polypeptide and Methods of Use Thereof
US7521230B2 (en) * 2002-10-16 2009-04-21 Board Of Regents Of The University Of Nebraska Nucleic acid encoding a Brain Derived Tau Kinase polypeptide and methods of use thereof
US7659446B2 (en) * 2003-02-25 2010-02-09 Mendel Biotechnology, Inc. Polynucleotides and polypeptides in plants
US7399587B2 (en) * 2003-05-29 2008-07-15 Astellas Pharma Inc. Canine CYP1A2 genetic polymorphism
US7507875B2 (en) * 2003-06-06 2009-03-24 Arborgen, Llc Transcription factors
US6939701B2 (en) * 2003-08-08 2005-09-06 Novozymes, Inc. Polypeptides having oxaloacetate hydrolase activity and nucleic acids encoding same
US20100037352A1 (en) * 2003-09-30 2010-02-11 Ceres, Inc. Sequence-determined dna fragments and corresponding polypeptides encoded thereby
US20060107345A1 (en) * 2003-09-30 2006-05-18 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20090081210A1 (en) * 2003-12-04 2009-03-26 Vaccinex, Inc. Methods of Killing Tumor Cells by Targeting Internal Antigens Exposed on Apoptotic Tumor Cells
US20050158323A1 (en) * 2003-12-04 2005-07-21 Vaccinex, Inc. Methods of killing tumor cells by targeting internal antigens exposed on apoptotic tumor cells
US7750209B2 (en) * 2003-12-17 2010-07-06 International Flower Developments Proprietary Limited Method for producing yellow flower by controlling flavonoid synthetic pathway
US7538204B2 (en) * 2004-01-21 2009-05-26 Malaysian Palm Oil Board Recombinant enzyme and uses therefor
US20120159672A1 (en) * 2004-02-06 2012-06-21 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20060150283A1 (en) * 2004-02-13 2006-07-06 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20060048240A1 (en) * 2004-04-01 2006-03-02 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20100037355A1 (en) * 2004-04-01 2010-02-11 Ceres, Inc Sequence-determined dna fragments and corresponding polypeptides encoded thereby
US7525016B1 (en) * 2004-05-21 2009-04-28 Iowa State University Research Foundation, Inc. Identification of syn-copalyl diphosphate synthase
US7195901B1 (en) * 2004-06-03 2007-03-27 The United States Of America As Represented By The Secretary Of Agriculture Diacylglycerol acyltransferase and its use to preferentially incorporate fatty acids into diacylglycerol
US7396979B2 (en) * 2004-06-30 2008-07-08 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying plant characteristics and phenotypes
US7709611B2 (en) * 2004-08-04 2010-05-04 Amgen Inc. Antibodies to Dkk-1
US7750207B2 (en) * 2004-09-01 2010-07-06 Monsanto Technology Llc Zea mays ribulose bisphosphate carboxylase activase promoter
US20100083407A1 (en) * 2004-09-30 2010-04-01 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying plant characteristics
US7569389B2 (en) * 2004-09-30 2009-08-04 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying plant characteristics
US7910800B2 (en) * 2005-08-15 2011-03-22 Evogene Ltd. Methods of increasing abiotic stress tolerance and/or biomass in plants and plants generated thereby
US7989676B2 (en) * 2006-08-31 2011-08-02 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics
US20100162433A1 (en) * 2006-10-27 2010-06-24 Mclaren James Plants with improved nitrogen utilization and stress tolerance
US20100011463A1 (en) * 2007-02-06 2010-01-14 Basf Plant Science Gmbh Compositions and Methods Using RNA Interference for Control of Nematodes
US20100017910A1 (en) * 2007-02-09 2010-01-21 Basf Plant Science Gmbh Compositions and Methods Using RNA Interference of CDPK-Like For Control of Nematodes
US20110105347A1 (en) * 2007-05-17 2011-05-05 Monsanto Technology Llc Corn polymorphisms and methods of genotyping
US20110167514A1 (en) * 2007-07-05 2011-07-07 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics
US20110179501A1 (en) * 2007-07-31 2011-07-21 The Ohio State University Research Foundation Methods for Reverting Methylation by Targeting DNMT3A and DNMT3B
US20090208564A1 (en) * 2007-08-27 2009-08-20 Chiang Jia Li Compositions of asymmetric interfering RNA and uses thereof
US20090094717A1 (en) * 2007-10-03 2009-04-09 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics
US20110008322A1 (en) * 2007-12-26 2011-01-13 Vaccinex, Inc. Anti-c35 antibody combination therapies and methods
US20120017338A1 (en) * 2008-01-15 2012-01-19 Wei Wu Isolated novel nucleic acid and protein molecules from corn and methods of using those molecules to generate transgenic plant with enhanced agronomic traits
US20110126316A1 (en) * 2008-05-01 2011-05-26 Academia Sinica Use of rice polypeptides/nucleic acids for plant improvement
US20110184045A1 (en) * 2008-06-30 2011-07-28 Gunther Hartmann Silencng and rig-i activation by dual function oligonucleotides
US20110177228A1 (en) * 2008-08-13 2011-07-21 Ceres, Inc. Plant nucelotide sequences and corresponding polypeptides
US20110145946A1 (en) * 2008-08-18 2011-06-16 Evogene Ltd. Isolated polypeptides and polynucleotides useful for increasing nitrogen use efficiency, abiotic stress tolerance, yield and biomass in plants
US20100168205A1 (en) * 2008-10-23 2010-07-01 Alnylam Pharmaceuticals, Inc. Methods and Compositions for Prevention or Treatment of RSV Infection Using Modified Duplex RNA Molecules
US20120017292A1 (en) * 2009-01-16 2012-01-19 Kovalic David K Isolated novel nucleic acid and protein molecules from corn and methods of using those molecules to generate transgene plants with enhanced agronomic traits
US20120023611A1 (en) * 2009-02-27 2012-01-26 Yongwei Cao Isolated Novel Nucleic Acid and Protein Molecules from Corn and Methods of Using Thereof
US20110201667A1 (en) * 2009-07-20 2011-08-18 Protiva Biotherapeutics, Inc. Compositions and methods for silencing ebola virus gene expression

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130313918A1 (en) * 2011-02-02 2013-11-28 Siemens Aktiengesellschaft Method for protecting charging cable, and charging device
US9524838B2 (en) * 2011-02-02 2016-12-20 Siemens Aktiengesellschaft Method for protecting charging cable, and charging device
CN104711260A (en) * 2013-12-13 2015-06-17 华中农业大学 Promoter Y8A for specific induced expression by recovering nitrogen supply after nitrogen deficiency of rice and application thereof
CN108410888A (en) * 2018-03-15 2018-08-17 山东农业大学 A kind of apple MdCEPR1 genes and its preparation method and application

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