KR100803390B1 - Root specific promoter - Google Patents

Abstract

A root specific promoter is provided to be able to be effectively used for mass-producing useful proteins in a transgenic storage root tissue and controlling the metabolism of a storage root tissue and the production of a functional material using a transformant by root-specifically inducing a target protein gene such as genetically modified proteins or proteins introduced by a foreign useful gene. A nucleoside phosphatase promoter includes a sequence of SEQ ID : NO. 1 and root-specifically expresses. A primer for amplifying the nucleoside phosphatase promoter consists of sequences of SEQ ID : NOs. 6 and 7. A foreign protein expression vector is characterized in that the nucleoside phosphatase promoter is introduced and further comprises a bar of SEQ ID : NO. 16 or an Egfp:gus of SEQ ID : NO. 17. A transgenic plant is characterized in that the foreign protein expression vector is transformed.

Description

Root specific promoter

1 shows genes AtNP and genes encoding endogenous actin (actin) genes, which encode Arabidopsis nucleoside phosphatase after reverse transcription using total RNA extracted from Arabidopsis root, leaf, stem, flower and seed tissue, respectively. Electrophoresis pictures showing gene bands amplified with primers that can be specifically amplified.

<Description of the code>

AtNP-Arabidopsis nucleoside phosphatase gene

actin -Arabidopsis-actin gene

FIG. 2 is a schematic diagram of the gene maps of the plasmids pBGWFS7-P AtNP (FIG. 2A) and pBGWFS7-P35S (FIG. 2B).

<Description of the code>

LB-Left border

Tnos-nos Terminator

BAR -herbicide resistance gene

Pnos-nos promoter

PAtNP-nucleoside phosphatase promoter

P35S-Cauliflower Virus 35S Promoter

EGfp-green fluorescent protein gene

GUS -β-glucuronidase Gene

T35S-Cauliflower Virus 35S Terminator

RB-Right border

Figure 3 is specifically using the total RNA extracted from each tissue of the Arabidopsis transformed with pBGWFS7-PAtNP specifically after the reverse transcription reaction Egfp enhanced and actin gene (actin), the Arabidopsis endogenous gene as a positive control Electrophoresis picture showing gene band amplified with primers to amplify.

<Description of the code>

M-1kb ladder DNA marker

GFP-Green Fluorescent Protein Gene

EF1a-Elongation factor 1a

Figure 4 shows the blue color reaction during seedling and leaf development of Arabidopsis transformed with pBGWFS7-PAtNP and pBGWFS7-P35S.

<Description of the code>

Transgenic Arabidopsis Transgenic Arabidopsis of PAtNP :: GUS-pBGWFS7-PAtNP

P35S :: GUS-pBGWFS7-P35S Transgenic Arabidopsis Plant

col0-nontransforming plant

Week 1-Baby pole seedlings 1 week after germination

2 weeks-Arabidopsis plants 2 weeks after germination

4 weeks-Arabidopsis plants 4 weeks after germination

5 is a green fluorescence image observed in the root cells of Arabidopsis transformed with pBGWFS7-P AtNP . FIG. 5A shows each detailed histology of the root, and FIG. 5B shows a green fluorescence image observed in the entourage site.

<Description of the code>

GFP-green fluorescent microscope image

Light-General Fluorescence Microscope Image

Merged-Merged image of green and green fluorescent images

The present invention relates to a root tissue specific promoter that regulates by restricting the expression site of a foreign useful gene to the root in the plant, and more particularly, the nucleoside phosphatase promoter of SEQ ID NO: 1 isolated from Arabidopsis; It relates to a primer, a transformation vector into which the promoter is introduced, and a transgenic plant transformed thereto.

Crop molecular breeding is the key to lead the next generation of agriculture in that it is possible to use all kinds of genes as a material and to fine-tune the effects of infinite breeding by pulling down the breeding technology that was possible only in the existing genome. Technology.

In order to maximize the effects of these crop molecular breeding techniques, 1) accumulation of a large amount of genetic data that can represent genes of various plants must be preceded, 2) various crop transformation systems must be established, and 3) external Development of various promoters capable of controlling the expression of the inserted gene should be preceded.

Known promoters used to achieve transformation purposes by confining the expression of foreign genes to specific tissues of plants can be classified as follows according to their function.

First, it is a root tissue specific expression promoter. Although no commercialized case yet, Arabidopsis peroxidase (peroxidase, prxEa) was isolated to confirm root tissue specific expression. Recently, the sweet potato-derived Maz gene ( ibMADS ) and sugar-induced ADP-glucose pyrophosphatase (ADP- Glucose pyrophosphatase (AGPase) gene was isolated and the promoter induces specific expression in roots and has been confirmed to induce root-specific transient expression in carrots and radishes (Korean Patent Publication Nos. 10-18771 and 10-604191). ). These promoters can be expected to be used primarily for the purpose of improving agricultural traits, accumulating useful proteins and producing useful substances in major root crops used for food, food, or food.

Second, seed specific promoters can be mentioned. As a representative example, the rice gluten promoter used to develop golden rice as a promoter of the major storage protein gene of rice has been widely used to induce seed-specific expression of monocotyledonous plants. Promoters mainly used to induce seed specific expression include soybean-derived lectin promoters, cabbage-derived napin promoters, and gamma-tocopherol methyl transferase (γ-TMT) in seedlings of Arabidopsis. Carrot-derived DC-3 promoters used in studies that enhanced the production of vitamin E by inducing gene expression, and there is a case confirmed the seed specific expression of the perilla-derived oleosin (oleosin) promoter. The seed specific promoters are mainly used for the purpose of accumulating useful proteins and producing useful substances in major crops in which the seeds themselves are used as food, food or food ingredients.

Third, a systemic expression induction promoter may be mentioned. The promoter of 35S RNA gene of CaMV (caaflower mosaic virus) is used as a representative dicotyledonous promoter as a plant whole-body induction promoter. ) And corn ubiquithin gene promoters have been mainly used, and recently, a promoter of rice cytochrome C gene (OsOc1) has been developed and used by domestic researchers (Korean Patent Publication No. 10-429335). These are already inherent to induce the expression of antibiotic or herbicide resistance genes and reporter genes used as selection markers in the basic plant transgenic carriers. Promoters are considered.

Fourth, other tissue specific promoters, such as a leaf, can be mentioned. Induced expression of potent genes only in photosynthetic tissues such as leaves.RBCS (ribulose bisphosphate carboxylase / oxygenase small subunit) promoter from rice and corn, RolD promoter to induce plant root expression from Agrobacterium. Patatin promoter, tomato-derived fruit maturation-specific expression-induced phytoene synthase (PDS) promoter, and cabbage-derived oleosin (oleosin) promoter whose function has been confirmed to be pollen-specific in Chinese cabbage. The male sterile crop was developed by inducing the expression of the Bt protein gene, which is toxic to cells, by the carpet tissue-specific BcA9 promoter of the male organ organ of flowers (Korean Patent Publication No. 10-435143).

In addition, a large number of promoters have been reported for each tissue of various plant-derived plants for various purposes, and are still under development, but most of the promoters that have been commercialized or widely used among plant researchers are mentioned above. This belongs, and there is a need for further development of new promoters that can precisely regulate the site of gene expression according to the intention of the developer.

Therefore, the present inventors have studied the root tissue specific promoter that regulates by restricting the expression site of the foreign useful gene in the plant, and confirmed that the nucleoside phosphatase promoter isolated from Arabidopsis is specifically expressed in the root tissue. This invention was completed.

Accordingly, an object of the present invention is to provide a nucleoside phosphatase promoter of SEQ ID NO: 1, a primer for amplifying it, a transformation vector into which the promoter is introduced, and a transformed plant transformed thereto.

In order to achieve the above object, in the present invention, nucleoside, which is a root tissue specific promoter of SEQ ID NO: 1, can be used when altering plant root components or improving agricultural characteristics by generating or accumulating foreign useful proteins and substances in plant roots. Phosphatase promoter P AtNP .

The present invention also provides a primer for amplifying the nucleoside phosphatase promoter P AtNP consisting of the bases of SEQ ID NO: 6 and SEQ ID NO: 7 for amplifying the nucleoside phosphatase promoter of SEQ ID NO: 1.

In addition, the present invention provides a foreign protein expression vector pBGWFS7-P AtNP comprising the nucleoside phosphatase promoter P AtNP of SEQ ID NO: 1.

The present invention also provides a transgenic plant cell transformed with the foreign protein expression vector pBGWFS7-P AtNP comprising the nucleoside phosphatase promoter P AtNP of SEQ ID NO: 1.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a root tissue specific promoter which regulates by restricting the expression site of a foreign useful gene to the root in a plant.

The present invention includes a nucleoside phosphatase promoter P AtNP (hereinafter referred to as "P AtNP ") which has a nucleotide sequence of SEQ ID NO: 1 and expresses root-specifically. Root tissue specific promoters according to the present invention are promoters involved in the transcription of nucleoside phosphatase protein genes isolated from Arabidopsis. The nucleoside phosphatase is generally known as a protein involved in cleavage of a phosphate group, which is one of the nucleic acid components, but the results of the gene function seen in Arabidopsis have not been reported yet. Annotation resulted in the nucleoside phosphatase protein gene. The promoter of the present invention is a promoter corresponding to 1,850 bp upstream of the nucleoside phosphatase gene coding site involved in the transcription of the nucleoside phosphatase gene that is specifically expressed only in root tissue.

The present invention includes a primer for amplifying the nucleoside phosphatase promoter P AtNP consisting of the sequences of SEQ ID NO: 6 and SEQ ID NO: 7. The primer is composed of a forward primer and a reverse primer each consisting of 32bp which specifically amplifies P AtNP , a root tissue specific promoter. The primer may be prepared by a method for synthesizing oligonucleotides known to those skilled in the art, and may be used by commercially synthesizing.

According to the present invention, a mutant based on the nucleotide sequence represented by SEQ ID NO: 6 and SEQ ID NO: 7 or a complementary nucleotide sequence thereof is partially deleted from an nucleotide sequence, addition of an excess base or nucleotide sequence, or a base or partial sequence of a nucleotide sequence in a nucleotide sequence. Substitution with other sequences, or primers consisting of their complex sequences may also be included.

The present invention includes a foreign protein expression vector pBGWFS7-P AtNP comprising the nucleoside phosphatase promoter P AtNP of SEQ ID NO: 1. The expression vector pBGWFS7 AtNP-P as well as the P AtNP of SEQ ID NO: 1 according to the present invention, the herbicide resistance gene SEQ ID NO: 16 of the bar or the reporter gene in SEQ ID NO: 17 of Egfp: may include gus.

The present invention includes a transgenic plant transformed with a foreign protein expression vector pBGWFS7-P AtNP comprising P AtNP of SEQ ID NO: 1. Plants to be transformed in the present invention include root crops such as radish, turnip, cabbage, cabbage, carrot, burdock, sweet potato, hemp, lotus root, potato and ginger; Medicinal plants such as ginseng, 칡 and deodeok; Feed crops such as corn and soybeans; Main grain crops such as rice; Plants that can be used as a root, including oilseed crops, such as rapeseed, but not limited to providing a transgenic plant that is asexual growth by tissue culture.

Examples of the target gene expressed by the transcriptional activity of the root tissue specific promoter of the present invention may be genes related to disease resistance or disease defense signaling for root disease control for agricultural transformation of crops to be transformed. In particular, in the case of medicinal root crops such as ginseng, introducing a pathogenic gene can also be expected to reduce the amount of pesticide control pesticides that are most problematic when cultivating for years. In addition, if you want to enhance specific nutritional ingredients, such as tocopherol and beta-carotene, at the root of crops for the purpose of producing or enhancing high value-added medicinal ingredients or specific active ingredients using cultured root cells of root crops, Biosynthetic genes or biosynthetic genes related to medicinal components such as saponin, gamma linolenic acid, and the like, but are not limited thereto, and may include various target genes that may be regulated using the root tissue specific promoter of the present invention.

The present invention may be understood in more detail by the following examples, which are intended to illustrate the present invention and are not intended to limit the protection scope of the present invention. On the other hand, commonly used DNA test methods not specifically mentioned in the following examples, see Sambrook et al. "Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, second edition, 1989".

Example 1 Expression Analysis of Arabidopsis Nucleoside Phosphatase Gene

To investigate the expression pattern of Arabidopsis nucleoside phosphatase gene, total RNA was isolated from pods including Arabidopsis root, leaf, stem and flower and seed during development. reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was performed.

In more detail, the total RNA was separated, and about 1 g of each sample was crushed into a mortar using a liquid nitrogen, transferred to a 2 ml tube, and 500 µl of RNA extraction buffer (50 mM sodium acetate pH 5.5, 150 mM LiCl, 5 mM EDTA, 0.5% SDS) and 500 µl phenol were added and mixed. The mixed solution was treated at 65 ° C. for 10 minutes and then mixed using a stirrer (rotary shaker) at room temperature for 15 minutes. After centrifugation at 4 ° C (10000rpm, 10 minutes), the supernatant layer was carefully transferred to a new tube, the mixture was mixed well by adding 500 μl of chloroform and centrifuged again to obtain the supernatant. 0.6 M volume of 8M lithium chloride (LiCl) was added thereto, and then stored at -20 ° C for at least 2 hours. After centrifugation at 4 ° C. and 12,000 rpm for 20 minutes, the RNA precipitate was washed once with 4M LiCl and 80% ethanol, and the final RNA precipitate was dissolved in 50 μl of distilled water. RNA eluate was quantified by measuring A ₂₆₀ / A ₂₈₀ using an UV spectrophotometer, and then treated with 50 μM of RNA as a template and 50 minutes at 50 ° C and 5 minutes at 85 ° C using dT primer. After cooling on ice, 1 μl of RNase H was added thereto, followed by treatment at 37 ° C. for 20 minutes to synthesize cDNA (10X RT buffer, 25 mM MgCl _2, 0.1M DTT, RNase OUT, SuperScript ^TM RT). CDNA as a template, the NP-F primer of SEQ ID NO: 2 (5'-TATTTGCGTGGCTATTGTTT-3 '), which is a set of nucleoside phosphatase gene AtNP specific primers, and the NP-R primer of SEQ ID NO: 3 (5'-CAATAATACAATCCCACAAACGAG-3') Sets were used.

In addition, the act-F primer of SEQ ID NO: 4 (5'-CATCAGGAAGGACTTGTACGG-3 ') and the act-R primer of SEQ ID NO: 5, which is a Arabidopsis actin primer set for demonstrating uniformity of the relative hem of the used RNA, 5'-GATGGACCTGACTCGTCATAC-3 ') was denatured at 95 ° C for 10 minutes, followed by 30 seconds of reaction at 95 ° C, 30 seconds of annealing at 65 ° C, and 30 times of polymerization at 72 ° C. Then PCR was performed at 72 ° C. for 7 minutes.

As a result of the reverse transcription PCR reaction, the transcript of the nucleoside phosphatase gene derived from Arabidopsis was not detected in pod tissues including leaves, stems, flowers, and seeds, but specifically detected in root tissues (FIG. 1). At this time, the relative amount of RNA used was demonstrated to be uniform using Arabidopsis actin specific primers. Thus, it was confirmed that the nucleoside phosphatase gene of Arabidopsis is specifically expressed in the root.

Example 2 Transformation Plasmid Preparation

1.85kb including a TATA box above a start codon of a nucleoside phosphatase gene (At1g14240) by searching for a gene corresponding to a nucleoside phosphatase specifically expressed in root tissues from the published Arabidopsis nucleotide sequence The region (SEQ ID NO: 1) was isolated and a P AtNP promoter function assay vehicle was constructed using the gateway carrier pBGWFS7 purchased from the University of Ghent, Belgium. A small amount of leaf tissue of the transgenic Arabidopsis plants was placed in an Eppendorf tube and separated and purified using a Genomic DNA Purification Kit (IJBIO DNA System). DNA eluate was quantified by measuring A ₂₆₀ / A ₂₈₀ using an ultraviolet spectrophotometer (UV spectrophotometer). Pnp-B1 primer of SEQ ID NO: 6 designed to specifically amplify the P AtNP promoter, including a portion of the recombinant nucleic acid sequence that the bacterium has specifically from the genome DNA isolated from the Arabidopsis bacteriophage (5'-AAAAAGCAGGCTAGGTGAATATCGTTCCTGTG-3 ') and Pnp-B2 primer (5'-AGAAAGCTGGGTGGATCGTTGCATTAACGTCT-3') set were prepared and the promoter site of 1.85 kb was regenerated through a PCR (polymerase chain reaction). Separated. At this time, PCR conditions were denatured at 95 ° C for 10 minutes, followed by 1 minute reaction at 94 ° C, followed by annealing reaction at 55 ° C for 1 minute, and a series of reactions for 30 minutes at 68 ° C for 2 minutes, followed by polymerization at 68 ° C for 10 minutes. The reaction was carried out.

At this time, in order to compare the function of the P AtNP promoter with the activity of the 35S promoter which is mainly used for transformation of a dicotyledonous plant, a sequence designed to amplify a 35S promoter of about 860bp while including a part of a bacterial-specific recombinant nucleic acid sequence site A 35S-F primer (5'-AAAAAGCAGGCTGGTCCCCAGATTAGCCT-3 ') of No. 8 and a 35S-R primer (5'-AGAAAGCTGGGTCCCGGGGATCCTCTAGA-3') of SEQ ID NO: 9 were prepared to isolate the 35S promoter site. The amplified PCR products contained adapter primer B1 (5'-GGGGACAAGTTTGTACAAAAAAGCAGGCT-3 ') and SEQ ID NO: 11 adapter primer B2 (5') containing the entire bacterial-specific recombinant nucleic acid sequence site when the bacteria were infected with bacteriophage. PCR amplification was again performed using -GGGGACCACTTTGTACAAGAAAGCTGGGT-3 '). The PCR reaction was denatured at 95 ° C. for 3 minutes, followed by 30 seconds at 94 ° C., followed by annealing reaction at 45 ° C. for 30 seconds, and a series of four reactions at 68 ° C. for 2 minutes, followed by 30 seconds at 94 ° C. Reaction, a series of reactions of 30 seconds at 55 ℃, polymerization for 2 minutes at 68 ℃ 19 was carried out by polymerization at 68 ℃ for 10 minutes.

The secondary PCR products were cloned by pDONR201 vector containing the bacteriophage-specific recombinant nucleic acid sequence site when the bacteria were infected with bacteriophage to produce a cloning vector of pDONR-P AtNP and pDONR-P35S. These two subcloned carriers were then subjected to an LR recombination reaction with the pBGWFS7 vector incorporating the reporter system linked to the Egfp and Gus genes to produce the final plant transformation carrier, followed by the final pBGWFS7-P AtNP and pBGWFS7- The P35S vehicle was completed (FIGS. 2A and 2B).

At this time, pBGWFS7 used as a binary vector has a spectinomycin resistance gene for microbial selection, and contains a herbicide resistant Bar gene as a plant selection factor. The transforming carriers were transformed into Agrobacterium tumefaciens strain GV3101 according to the direct Agrobacterium transformation method (see Holster et al., Freeze-thaw method, 1978) using liquid nitrogen, followed by an alkali method such as An. The introduction into Agrobacteria was confirmed using an Agrobacterium quick-screen based on alkaline lysis.

Example 3 Plant Material and Arabidopsis Transformation

After seeding, primary harvesting of Arabidopsis thaliana ecotype Columbia plants grown for about 4 weeks in a 23 ° C incubator (16 hours day / 8 hours night) was removed to induce at least 3-4 secondary harvests. Agrobacteria were infected by spraying the Agrobacteria cultures containing the conversion carriers with a suspension of OD 0.8 diluted in a solution containing 5% sucrose and 0.05% silwet77 to the flower buds. After overnight incubation with sufficient humidity and dark conditions, incubate for 4-5 days in a 23 ° C incubator (16 hours day / 8 hours night), and then culture and dilute Agrobacteria in the same manner as above. Incubated overnight with sufficient humidity and dark conditions, and then grown for 3 to 5 weeks in a 23 ° C. incubator (16 hours day / 8 hours night) until the lysics were fully browned. Then harvested. The seeded seeds were sown again and the transformants of T1 generation were selected by spraying 0.3% batha on the second and third weeks of growth. T1 transgenic plants were grown for about 2 months in a 23 ° C. incubator (16 hours day, 8 hours night) until the pod tissue (silique) had fully matured to brown, followed by T2 seeding.

Example 4 RT-PCR Reaction of Transgenic Plants

Sowing of transgenic T2 seeds and spraying 0.3% batha on the second and third weeks of growth, leaves, stems, roots, flowers and seed pods of the transgenic Arabidopsis plants (including seeds at 3 weeks after flowering) About 5 ㎍ of total RNA extracted from various tissues in the same manner as in Example 1 was treated with 50 uM oligo dT primer for 50 minutes at 50 ° C. and 5 minutes at 85 ° C., and then cooled on ice, and then 1 μl of RNase H was added thereto. CDNA was synthesized by treatment at 37 ° C. for 20 minutes (10X RT buffer, 25 mM MgCl _2, 0.1M DTT, RNase OUT, SuperScript ^TM RT), and the synthesized cDNA was used as a template. Arabidopsis extension to demonstrate uniformity of the relative amounts of RNA used and GFP-F primer of 12 (5'-CGACAAGCAGAAGAACGGCATCAA-3 ') and GFP-R primer of SEQ ID NO: 13 (5'-TGTAACGCGCTTTCCCACCAACTC-3') Elongation factor (EF-1α) -1α) Denatured at 95 ° C. using EF1α-F primer (5'-GTTCACATTAACATTGTGGTCATT-3 ') of SEQ ID NO: 14 and EF1α-R primer (5'-CAGGTACCAGTGATCATGTTCTTG-3') of SEQ ID NO: 15 for 10 minutes Thereafter, the reaction was carried out at 95 ° C. for 30 seconds, followed by annealing reaction at 55 ° C. for 30 seconds, and a series of 35 reactions at 72 ° C. for 30 seconds, followed by PCR at 72 ° C. for 7 minutes.

The results of the RT-PCR reaction, P AtNP promoter, whereas transgenic aneunde not detected body green fluorescent protein transcription of the gene Gfp in Arabidopsis leaves, stems, flowers and seed pod tissues, P AtNP promoter transgenic root tissues of Arabidopsis thaliana plants Only the Gfp transcript was clearly detected (FIG. 3). At this time, the relative amount of RNA used was demonstrated to be uniform using the Arabidopsis elongation factor EF-1α.

Example 5 GUS blue color reaction analysis

PAtNP Promoter and 35S promoter transgenic T2 seeds were sterilized in 25% lacx for 7 minutes, and then plant-growing MS medium containing 0.75 mg / L of phosphinotricin, a buster herbicidal component, for selection of transformants. Germinated in As a control, untransformed Arabidopsis plants were germinated in MS medium without phosphinothricin. Transgenic Arabidopsis plants at 1, 2 and 4 weeks after germination were assayed for GUS-assay buffer [Sol. I: 20 g of cyslohexyl ammonium salt (X-gluc), Sol. II: NaH ₂ PO ₄ H ₂ O 100mM, NaEDTA 10mM, Triton-X-100 0.1%, pH7.5] and treated at 37 ° C. for 24 hours, and then treated with 70% ethanol to remove chlorophyll. . Plants were observed using a light microscope or visually, and the results are shown in FIG. 4. PAtNP from seedling stage of the first week of germination in the results shown in Figure 4 Promoter transformants showed GUS blue color specific to the root. GUS blue coloration was not observed in the nontransgenic Arabidopsis plants at 2 and 4 weeks, and 35 US promoter transforms were observed in whole body plants, while GUS blue coloration was observed in whole body plants. In promoter transgenic plants, GUS blue color was observed only in the roots.

Example 6 GFP Green Fluorescence Image Analysis

PAtNP Promoter T2 generation seeds in 25% lacx for 7 minutes and germinate in plant growth MS medium containing phosphinotricin at a concentration of 0.75 mg / L to prepare 9-day-seedling. It was. Confocal microscopy was used to capture GFP fluorescence. Green fluorescence images were observed on seedlings 9 days after germination, and PAtNP promoter showed strong fluorescence expression in newly formed root hairs of root tissues. As a result of observing the blue color reaction by the method described in Example 5 of the same material, blue color was also confirmed in the newly formed root hair tissue.

Based on the above experimental results, it was confirmed that the P AtNP promoter induces the expression of the root tissue specific expression of the reporter gene through the transcriptional step and the blue color reaction and the green fluorescence image by the final reporter protein function. (FIGS. 5A and 5B).

As described above, the root tissue specific promoter of SEQ ID NO: 1 according to the present invention affects plant growth by inducing root-specifically a target protein gene such as a protein changed by genetic engineering method or a protein by foreign useful gene introduction. It is effective to limit the expression to the roots and to effectively produce a large amount of useful protein in transformed storage root tissues or to control metabolism and production of functional substances in transformed storage root tissues using transformants. Could.

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Claims (6)

A nucleoside phosphatase promoter having a nucleotide sequence of SEQ ID NO: 1 and expressing root-specifically. A primer consisting of the sequences of SEQ ID NO: 6 and SEQ ID NO: 7 and for amplifying the nucleoside phosphatase promoter according to claim 1. A foreign protein expression vector into which the nucleoside phosphatase promoter of SEQ ID NO: 1 is introduced. The expression vector of claim 3, wherein the expression vector further comprises bar of SEQ ID NO: 16, which is a herbicide resistance gene, or Egfp: gus , SEQ ID NO: 17, of a reporter gene. A transgenic plant transformed with the foreign protein expression vector according to claim 3. The plant of claim 5, wherein the plant to be transformed comprises root crops including radish, turnip, cabbage, cabbage, carrot, burdock, sweet potato, hemp, lotus root, potato and ginger; Medicinal plants, including ginseng, 칡 and deodeok; Feed crops including corn and soybeans; Main crops including rice; And oilseed crops including rapeseed; transgenic plant, characterized in that selected from the group consisting of.

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