KR101444215B1 - BrSTY1 PROMOTER FROM BRASSICA AND PLANT TRANSFORMED WITH THE SAME - Google Patents

Abstract

The present invention relates to a BrSTY1 promoter that specifically induces expression in stem apical meristems, leaves, flowers and pods, comprising the nucleotide sequence of SEQ ID NO: 1. More particularly, the present invention relates to a promoter that specifically induces expression in truncated mesenchymal tissue, leaves, flowers, and pods, and a recombinant expression vector comprising the same, and a plant transformed with the recombinant expression vector. Since the BrSTY1 promoter of the present invention specifically induces expression in truncated mesenchymal tissues, leaves, flowers and pods, the expression of the target protein can be specifically expressed using the promoter, thereby improving the trait of the crop.

Description

The BrastY-derived BrSTY1 promoter and the plant transformed with the BrSTY1 promoter (BrSTY1 PROMOTER FROM BRASSICA AND PLANT TRANSFORMED WITH THE SAME)

The present invention relates to a Chinese cabbage-derived BrSTY1 promoter that specifically induces expression in stem apical meristems, leaves, flowers and pods. More particularly, the present invention relates to a promoter that specifically induces expression in truncated mesenchymal tissue, leaves, flowers, and pods, and a recombinant expression vector comprising the same, and a plant transformed with the recombinant expression vector.

Recently, as the genetic engineering technology has developed, many researches have been carried out to improve the characteristics of plants. In particular, there is much interest in techniques for obtaining a useful gene from a transgenic plant. A promoter involved in expression of the gene is required when the desired gene is expressed in a transgenic plant.

 In general, a promoter is a kind of regulatory region having a function of regulating the expression of a nucleotide sequence, that is, a gene, which determines when and where a gene is to be expressed. Over the years, numerous types of promoters have been identified through endless research. Conventionally reported promoters can be classified according to their features and functions such as a promoter that induces expression at all times and a promoter that induces expression in time or position specific manner.

More specifically, the first is a strong promoter capable of maximizing the expression level. This was initially developed as a promoter that is used to maximize the expression level of the target gene, and is representative of the 35S RNA gene promoter (P35S) of flower cabbage mosaic virus and the actin gene promoter of rice. They are mainly used for the purpose of production of antibiotic resistance genes used as selection markers in the transformation of plants, genes for inhibiting insect pests of plants, or plants.

Second, promoters that limit the time of expression can be mentioned. This has been reported as a promoter that restricts the expression of a gene of interest only to a specific stage of the developmental stage of the plant. For example, promoters of genes involved in flowering can express genes only during the flowering season, and promoters of genes expressed by various biotic or abiotic stimuli can be included in the timing-regulated promoters.

Third, tissue-specific promoters that limit expressed tissue are examples. This is the promoters that are used to achieve the transfection purpose by limiting the expression of the desired gene only to specific tissues of the plant. A number of examples of promoters inducing such tissue-specific expression have been reported for each tissue of a plant according to various purposes to be used.

In particular, the promoters inducing tissue-specific expression among the promoters classified according to functions and characteristics are classified as follows according to the tissues expressed in the plant. First, a systemic expression-inducible promoter can be mentioned. As a promoter that induces expression in plant whole body, a promoter of 35S RNA gene of cauliflower mosaic virus (CaMV), which is also a promoter used to maximize the expression level, is used as a representative promoter for a dicotyledonous plant. Promoters of rice actin and maize ubiquitin genes have been mainly used as promoters for systemic expression of the plant in the monocotyledonous plant. Recently, a promoter of rice cytochrome C gene (OsOc1) has been developed 0429335). These genes are used for the purpose of inducing the expression of antibiotics, herbicide resistance genes and reporter genes used as selection markers in the transformation as in the case of the promoters maximizing the expression level. From the viewpoint of research, These are the promoters that are considered to be the priority when attempting to reveal.

Second, seed-specific promoters can be mentioned. As a representative example, the rice glutelin promoter used for the development of golden rice as promoters of rice major storage protein gene is widely used to induce seed-specific expression of monocotyledonous plants. Promoters that are mainly used to induce seed-specific expression include soybean-derived lectin promoter, cabbage-derived napin promoter and γ-tocopherol methyl transferase (γ-TMT) in Arabidopsis seeds. There are carrot-derived DC-3 promoter and perilla derived oleosin promoter used in studies to promote vitamin E production by inducing gene expression.

Third, root-specific expression promoter has not yet been commercialized. However, root-specific expression of pyruvate peroxidase (prxEa) has been isolated and confirmed recently. In recent years, the expression of maz genes (ibMADS) It has been reported that the ADP-glucose pyrophosphatase (AGPase) gene is isolated and the corresponding promoter induces a specific expression in the root and induces root-specific transient expression in carrot and radish .

Fourthly, there are other tissue specific promoters such as leaves. The related promoters include ribosyl bisphosphate carboxylase / oxygenase small subunit (rbcS), which induces expression of strong genes only in photosynthetic tissues of leaves and the like. ) Promoter, tomato-derived fruit maturation-specific expression-inducing phytoene synthase (PDS) promoter, and pollen-specific plant-derived cabbage-derived oleosin promoter.

Such seed, root or leaf-specific expression promoters are expected to be used for the purpose of improving agricultural traits, accumulating useful proteins, and producing useful substances in major crops used for food, food, or food.

However, conventional promoters known to induce tissue-specific expression in the known promoters, particularly plants, have the advantage of being capable of expressing a target gene in a tissue-specific manner, but have a disadvantage in that the expression amount thereof is insignificant. Therefore, it is urgent to find useful new promoters capable of expressing a gene of interest in a plant in a tissue-specific and large-scale expression.

Accordingly, the present inventors have continued research on a novel promoter capable of expressing a target gene in a tissue-specific manner in a plant, and have found that a promoter isolated from Chinese cabbage has an activity of inducing the expression of a target gene in a specific tissue at the time of plant growth and development And completed the present invention.

Korean Patent No. 10-1054891

It is an object of the present invention to provide a BrSTY1 promoter that specifically induces expression in stem apical meristem, leaf, flower, and pod, comprising the nucleotide sequence of SEQ ID NO: 1.

To provide a recombinant expression vector comprising a foreign gene encoding a protein of interest operably linked to the promoter of the present invention.

It is still another object of the present invention to provide a microorganism transformed with the recombinant expression vector and a plant transformed with the transformed microorganism.

It is still another object of the present invention to provide a method for producing a transgenic plant in which a target protein is specifically expressed in truncated mesenchymal tissues, leaves, flowers and pods using the promoter.

The present invention provides a BrSTY1 promoter that specifically induces expression in stem apical meristems, leaves, flowers and pods, comprising the nucleotide sequence of SEQ ID NO: 1.

The leaves may be hydathodes, veins, midribs and trichomes of the leaves.

The invention also provides a recombinant expression vector comprising a foreign gene encoding a protein of interest operably linked to a promoter.

According to one embodiment of the present invention, the recombinant expression vector may be pPBrSTY1.

The present invention also provides a microorganism transformed with said recombinant expression vector.

According to an embodiment of the present invention, the microorganism may be Agrobacterium tumefaciens .

The present invention also provides a transgenic plant transformed with the above-mentioned transforming microorganism.

According to one embodiment of the present invention, the plant may be a Arabidopsis.

Further, according to the present invention,

Preparing a recombinant vector comprising a promoter consisting of the nucleotide sequence of SEQ ID NO: 1 and a foreign gene encoding a target protein operably linked thereto;

Transforming the microorganism with the recombinant expression vector to prepare a transformed microorganism; And

Infecting the plant with the transformed microorganism;

Wherein the target protein is expressed specifically in stem apical meristems, leaves, flowers and pods.

The present inventors isolated a promoter region (-2.1 kb) of the STY1 (STYLISH 1) gene, which is one of the SHI (SHORT INTERNODE) genes of Chinese cabbage, in order to discover a novel promoter that specifically expresses a gene of interest in plants, Respectively. Specifically, the gene specific expression of BrSTY1 (Brassica rapa STYLISH1) gene was identified and the BrSTY1 promoter region (- 2.1 kb) using Chinese cabbage BAC clone was isolated. The thus obtained DNA base sequence is shown in SEQ ID NO: 1 and named "BrSTY1 promoter" (see Example 1).

The present inventors confirmed the expression pattern of the GUS gene in the Arabidopsis thaliana by fusing the GUS gene to the BrSTY1 promoter of the present invention in order to confirm whether the BrSTY1 promoter of the present invention expresses the target gene in a tissue-specific manner See Examples 3-4). As a result, it was confirmed that the GUS gene was expressed in truncated mesenchymal tissues, leaves, flowers and pods of Arabidopsis thaliana (see Fig. 2-4).

From the above results, it can be seen that the BrSTY1 promoter of the present invention is a promoter which induces the expression of the target gene in truncated mesenchymal tissues, leaves, flowers and pods of plants.

The term "promoter" is a gene site located upstream of an encoding site, which generally includes a point at which transcription is initiated. The promoter is a TATA box that controls gene expression, a CAAT box site, And an enhancer that promotes the expression of almost all genes regardless of the site, position, and direction. In addition, among these promoters, there are promoters that induce expression constantly in all tissues and promoters that induce expression in a time or position specific manner. In particular, the promoter according to the present invention induces expression specifically in specific parts of plants do.

The "expression vector" refers to plasmids, viruses or other vectors known in the art in which the promoter of the present invention and the gene sequence encoding the target protein operably linked to the promoter can be inserted or introduced . The nucleotide sequence encoding the plant-specific expression promoter of the plant according to the invention and the target protein operably linked to the promoter may be operably linked to an expression control sequence, wherein the operably linked gene sequence and the expression control sequence May be included in an expression vector that also contains a selection marker and a replication origin.

Said "operably linked" may be a gene and expression control sequence linked in such a way as to enable gene expression when a suitable molecule is coupled to an expression control sequence. "Expression control sequence" means a DNA sequence that regulates the expression of a polynucleotide sequence operably linked to a particular host cell. Such regulatory sequences include promoters for conducting transcription, any operator sequences for regulating transcription, sequences encoding suitable mRNA ribosome binding sites, and sequences controlling the termination of transcription and translation.

The expression vector according to the present invention may be prepared by inserting a promoter of the present invention using a conventional vector used for protein expression as a basic framework and inserting a nucleotide sequence encoding a target protein downstream of the promoter have. Therefore, plasmids derived from Escherichia coli (pBR322, pBR325, pUC118 and pUC119), Bacillus subtilis -derived plasmids (pUB110 and pTP5), yeast-derived plasmids (YEp13, YEp24 and YCp50 ) And Ti plasmids, plant virus vectors, and binary vectors such as pCHF3, pPZP, pGA, and pCAMBIA sequences can be used. However, any person skilled in the art can use any vector as long as it can introduce the promoter of the present invention and the base sequence encoding the target protein operably linked to the promoter into the host cell.

The target protein that can be used in the present invention may be any target protein derived from the outside, that is, an exogenous protein or an endogenous protein and a reporter protein, which is desired to be expressed specifically by a plant tissue by the promoter of the present invention. The exogenous protein refers to a protein that does not naturally exist in a specific tissue or cell, and the endogenous protein refers to a protein expressed by a gene naturally present in a specific tissue or cell. In addition, the reporter protein is a protein expressed by a reporter gene, and refers to a marker protein that shows its activity in a cell by its presence.

Therefore, when the promoter of the present invention is used, in the case of a crop which consumes a specific part of a plant, for example, a leaf or a stem, a target protein can be taken together with a leaf or a stem by expressing a useful protein of interest in a specific tissue of the plant You can develop a crop.

In one embodiment of the present invention, the BrSTY1 promoter of the present invention having the nucleotide sequence of SEQ ID NO: 1 is operably linked to a known vector having a reporter system in which a blue coloring gene, Gus, One recombinant expression vector, pPBrSTY1, was prepared (see Examples).

A recombinant expression vector comprising a promoter of the present invention and a gene sequence encoding the target protein operably linked to the promoter may be introduced into a host cell using methods known in the art. Methods for introducing the recombinant expression vector according to the present invention into host cells include, but are not limited to, calcium chloride (CaCl 2) and heat shock method, particle gun bombardment, silicon carbide whisker (Silicon carbide whiskers), sonication, electroporation, and PEG (polyethylenglycol) precipitation.

Accordingly, the present invention provides a host cell transformed with a recombinant expression vector comprising a promoter of the present invention and a nucleotide sequence encoding a target protein operably linked to the promoter. The host cell is preferably a microorganism. In one embodiment of the present invention, Agrobacterium tumefaciens GV3101 was used.

In addition, the recombinant expression vector of the present invention can be introduced into plants using methods known in the art. For example, but not by way of limitation, Agrobacterium sp. -Mediated methods, particle gun bombardment, silicon carbide whiskers, sonication, Electroporation and precipitation by PEG (polyethylenglycol) can be used. Preferably, an Agrobacterium sp. -Mediated method can be used.

Accordingly, the present invention provides a transformed plant by introducing into the plant a recombinant expression vector comprising a promoter of the present invention and a nucleotide sequence encoding a target protein operably linked to the promoter.

The transformed plant according to the present invention can be obtained by a conventional method in the art, such as an ovine reproduction method or an aseptic reproduction method. More specifically, the plant of the present invention can be obtained through reproductive process, which is a process of producing seeds through the moisture process of flowers and propagating from the seeds. Also, the plant can be transformed with the recombinant expression vector according to the present invention, and then obtained by the conventional method for inducing callus, rooting, and soil purification, through a silent propagation method. That is, a fragment of a plant transformed with the recombinant expression vector of the present invention is pelleted in a suitable medium known in the art, and cultured under appropriate conditions to induce callus formation. When shoots are formed, the plant is transferred to a hormone-free medium Lt; / RTI > After about two weeks, the shoots are transferred to rooting medium to induce roots. The transformed plants according to the invention can be obtained by roots being induced and then transplanted into the soil for purification. Transgenic plants in the present invention may include whole plants as well as tissues, cells or seeds obtainable therefrom.

The present invention also provides a recombinant vector comprising a promoter of the invention and a foreign gene encoding a target protein operably linked thereto; Transforming the microorganism with the recombinant expression vector to prepare a transformed microorganism; And infecting the plant with the transformed microorganism; Wherein the target protein is expressed specifically in stem apical meristems, leaves, flowers and pods.

Since the promoter of the present invention specifically induces expression in stem apical meristems, leaves, flowers and pods, it is possible to improve the trait of a crop by specifically expressing a target protein using the promoter.

FIG. 1 shows a vector map for plant transformation according to an embodiment of the present invention. The reporter gene GUS is regulated by the BrSTY1 promoter and contains a hygromycin resistance gene as an antibiotic screening gene.
FIG. 2 is an analysis of the BrSTY1 promoter activity in the stages of growth development. 0d to 35d: Transgenic Arabidopsis thaliana shows the 35th day after germination in the seed germination stage.
FIG. 3 is a graph showing the results of GUS expression analysis of the transformed Arabidopsis thaliana into which the BrSTY1 gene promoter was introduced, according to an embodiment of the present invention. 35d: rosette leaf, Ht: hydathode, V: Vein, Mr: midrib, Cl: cauline leaf, ), Tc: trichome.
FIG. 4 is a graph showing the results of GUS expression analysis at the reproductive growth stage of the transgenic Arabidopsis thaliana strain into which the BrSTY1 gene promoter has been introduced, according to an embodiment of the present invention. 35d: 35 days after germination Mature transgenic plants, RS: reproductive stage, F: flower, Fb: flower bud, Sty: style, Sp: calyx sepal, Si: silique, Rc: receptacle,

The present invention will be described in more detail by way of examples. It should be noted, however, that the present invention is not limited by the following examples.

Example 1: Promoter region separation and base sequence analysis

Using the nucleotide sequence of the BAC (KBrB088I08) clone of the cucumber branch, a region expected to be a promoter region of -2.1 kb including the TATA box and the like was searched above the BrSTY1 gene initiation codon, and a gene promoter specific primer was prepared as follows.

BrSTY1-Pro-Forward: 5'-GGATCCTGTACATAAATGTGTGCATACTAGTAG-3 '(SEQ ID NO: 2)

BrSTY1-Pro-Reverse: 5'-AAGCTTTCACCGGCAACACCACCACAAC-3 '(SEQ ID NO: 3)

A promoter region was isolated by performing a polymerase chain reaction (PCR) from a BAC clone. At this time, the PCR conditions were denaturation at 95 ° C for 5 minutes, followed by 35 cycles of 1 cycle at 95 ° C for 1 minute, 55 ° C for 1 minute, and 72 ° C for 2 minutes, followed by reaction at 72 ° C for 10 minutes. Then, the nucleotide sequence of the DNA obtained in the PCR reaction was confirmed, and the BrSTY1 promoter (-2.114 kb) was isolated and identified as SEQ ID NO: 1.

Example 2: Construction of Transformation Vector and Transformation of Plants

The BrSTY1 promoter isolated in Example 1 was inserted into pGEM-T-easy, digested with BamHI and HindIII, and inserted into pCAMBIA1381 digested with BamHI and HindIII. A plant transformation vector pPBrSTY1 having a reporter gene GUS fused to the BrSTY1 promoter was prepared and introduced into Agrobacterium tumefaciens GV3101 (Fig. 1).

In order to introduce the constructed vector pPBrSTY1 into Arabidopsis thaliana, Agrobacterium tumefaciens GV3101 was repeatedly frozen and thawed twice, and then LB medium containing 50 mg / l of antibiotic cannabis, 10 mg / l of rifampicin and 40 mg / l of gentamicin . The introduction of the carrier in the GV3101 strain was confirmed by colony PCR, and the colonies in which the gene introduction was confirmed were used for Arabidopsis transformation. Arabidopsis thaliana ecotype Col-o was transformed into wild-type Arabidopsis thaliana ecotype Col-o. The Arabidopsis thaliana was first cultivated for about 4 weeks at 22 ℃ growth (16 hours / 8 hours). After removing the primary chiasm of 4 weeks old Arabidopsis plants, 3-4 secondary chests of at least 8-10 cm were induced. Agrobacterium whose vector introduction was confirmed using the transformed GV3101 strain was inoculated on LB medium containing 50 mg / L of cannabimycin and cultured at 28 DEG C for about 2 days. A suspension containing 5% sucrose and 0.05% silwet L-77 precipitated by centrifugation of fully grown Agrobacterium was diluted to an OD of about 0.6. The suspension thus prepared was sprayed on the buds of the Arabidopsis thaliana and cultured for 24 hours in a state of maintaining sufficient humidity and dark conditions, followed by further culturing for 3-4 days on the growth conditions of 22 ° C, 16 hours day and 8 hours night, And the transformation was repeated. Thereafter, the seeds were harvested after growth for about 4-6 weeks until the pods were completely matured on the growth at 22 ° C, 16 hours day and 8 hours night. The harvested seeds were inoculated on MS medium containing 30 μg / ml hygromycin to select transformants, and the expression pattern of the BrSTY1 promoter was analyzed by GUS staining from selected transformants.

Example 3: Analysis of BrSTY1 promoter activity at each stage of development of Arabidopsis thaliana

In order to confirm whether the promoter of BrSTY1 gene of Chinese cabbage induces gene expression specifically in the plant, the growth stage of Arabidopsis transformed with recombinant expression vector inserted with the promoter of BrSTY1 gene and the expression pattern of GUS gene in each tissue Respectively. Namely, four lines isolated from Arabidopsis thaliana transformed in Example 2 were germinated in MS medium to observe GUS (blue coloring gene) expression pattern at the development stage of seedling, GUS gene expression pattern was observed at various stages of growth stage of leaves, stems, and roots in the plants at 15 and 35 days after sowing. The germinated seedlings were immersed in a GUS-assay buffer (20 mM of cyslohexyl ammonium salt, 100 mM of NaH ₂ PO ₄ .H ₂ O, 10 mM of NaEDTA, and 1000.1% of Triton-X, pH 7.5) ℃ for 24 hours, chlorophyll was completely removed with 70% ethanol to compare GUS gene expression patterns.

As shown in FIG. 2, from day 1 after seed germination, GUS expression was strongly expressed in apical meristems including hypocotyl until 35 days.

As shown in FIGS. 2 and 3, the GUS expression by the BrSTY1 promoter at 35 days was strongly expressed in shoot apical region and Vein along the midrib of the leaf, (hydathode). As shown in Fig. 3, GUS expression was also confirmed in trichomes of leaves. In root tissue, GUS expression was observed in the primary root of the plant from the 15th day until the 35th day, but not in the other growth stages. As shown in Fig. 3, GUS expression was also confirmed in the drainage tissue of the cauline leaf. Therefore, it can be seen that the BrSTY1 promoter of the present invention is a specific promoter that induces gene expression specifically in tissue during the developmental developmental period.

Example 4: Analysis of BrSTY1 promoter activity in developmental stages such as transgenic Arabidopsis blooming and seed maturation

 In order to confirm whether the promoter of BrastY1 gene of Chinese cabbage induces the expression of the gene in developmental stages such as flowering and seed maturation of the plants, a flower bud, a flower, a pod The expression of GUS in each tissue including silique was analyzed. Thus, it can be seen that the BrSTY1 promoter is a tissue-specific promoter that induces expression at the reproductive organs flower and pod region.

<110> REPUBLIC OF KOREA <120> PROMOTER FROM BRASSICA AND PLANT TRANSFORMED WITH THE SAME <130> P120914 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 2114 <212> DNA <213> Brassica rapa STYLISH1 <400> 1 tgtacataaa tgtgtgcata ctagtagctt tatacactaa atttaagcca caggtgcaaa 60 tctgcaccta aattgatatc ttcaaaagat tatgggcata tggataatct gaacaatata 120 ataataataa taatagtaat caatgtattg ccatttttaa ataagtagtg acttatttag 180 tccacatgtt tatcttcttg ctgaggattt tcgaaacctt gatttaacta accctcattg 240 ttagtatatg cgctacctcg tccactaata attgttttga aaggtttgat taccaatgat 300 ctcacatttg ctaagccgtt agataaaaaa acagatctca catttgctaa aataaggggc 360 ttttgacaag gtttcgtact cagttctaga tcatggttag gaggtctaac taataggtca 420 cagaatatac aactatcttt atttaaaatc ataaaactat cttaggatag tcaaaaatag 480 atttgtgaca ccataagagc ttgtcattag taggaactaa atggggtttc ataatgtaaa 540 attaaaattg attggctatc tcaaatttga gaaacttata tgataaatta ttagactaag 600 aacactctaa tataaaaata aattttaaaa aacaagccaa aataaaacat agaaaaactt 660 aattttgagt tgttcatcta atatttaata attagctagt aaaatatgtt aatttttatt 720 agataattga gatacattta gttttaggac tgcctcagag caagagcatt agaggttcaa 780 gagggaggtt cacacgtttt acgagaaaaa aaaatattaa aaaaagcaaa agtgatgaac 840 tcaccccttc tcacctcttc tgcatgatac cctctctcct aaagttcatc actgtagcgc 900 gggccccacg acacgtggcg gcccgcgatt ggtcaatttt tttttttttt tttttttttt 960 aaaaaatcaa aatgaaaaaa aagaaaaact ttaataataa aaaattaaga aggtgaaccc 1020 caaagggggg gttcactgat gctcatgctc tcagtgctaa aaacagttaa agtttctaag 1080 aaaagaaata atagtgtaat gatataaata tatataaata ttatttaaat gaatgtcata 1140 tgtcatttga aagtctcacc agtattttaa aagttctcaa acgaaaacct ttcactattt 1200 tcacatgctt attattatta ttattattat tattattatt attagtttta aaatactaga 1260 tactagttct tttaaatttc tgggatgaac atgcttttcg attatcaagt ttgaaatttt 1320 tacgaataat atttttagaa ttcatgggat gatcatttga aaattttact cctttaagga 1380 taattagtta gttctttttt ttttataacg actagttagt tcattttaac aaaacaaaac 1440 atataaaaat ttgcaaaatg taagagaaaa ggggaaagaa acaacgggtt tccgggaaga 1500 aacgacaaag aagaaaaata ccattggtgc cgacgtgaca aaacaatagc cgttggatca 1560 taaacgatca acgataagag attaatgttc tagcgggtgt ggcaattttg aagtgactag 1620 tgagaggtgt aaaacaaaaa taaagaaaaa agatgttttg caaaatctag aaaacaaagg 1680 aaaagagagg aggagacaga gaattaaatt agaggagcag agtggagaga gagatattaa 1740 ggttatggcg gcgttgcaga gcgcgagaaa ggccatgaaa ccctctctct gatctctctc 1800 tttcttcccc ttcttctttc tctctgtgtg tttcttaaat actctctctt ctctcgtacc 1860 gacaaacgag cttgcctttt taccacaccc ttaaaagctg tactcaactt caatttcaat 1920 aacactacaa acccggagag aatgaactct ctacctaaac cctagttttc ctctctcccc 1980 tatagtctca aagatctaag cgagaaagtt aaaaaagatc tagagcgaga gtctttttgt 2040 ttcatttttt ttcttctaaa tatctgattg gagagcggtg gtatgggtag tggttgtggt 2100 ggtgttgccg gtga 2114 <210> 2 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> BrSTY1-Pro-Forward Primer <400> 2 ggatcctgta cataaatgtg tgcatactag tag 33 <210> 3 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> BrSTY1-Pro-R Primer <400> 3 aagctttcac cggcaacacc accacaac 28

Claims (11)

A BrSTY1 promoter that specifically induces expression in stem apical meristems, leaves, flowers and pods, comprising the nucleotide sequence of SEQ ID NO: 1.
The method according to claim 1,
The BrSTY1 promoter wherein the leaves are hydathodes, veins, midribs and trichomes of leaves.
3. A recombinant expression vector comprising a foreign gene operably linked to the promoter of claim 1 or 2, encoding a protein of interest.
The method of claim 3,
Wherein said recombinant expression vector is pPBrSTY1 and has a cleavage map as set forth in FIG.
A microorganism transformed with the recombinant expression vector of claim 3. 6. The method of claim 5,
When the microorganism is Agrobacterium tumefaciens ).
A transgenic plant transformed with the transformed microorganism of claim 5.
8. The method of claim 7,
Wherein the plant is a transgenic plant.
Preparing a recombinant vector comprising a promoter consisting of the nucleotide sequence of SEQ ID NO: 1 and a foreign gene encoding a target protein operably linked thereto;
Transforming the microorganism with the recombinant expression vector to prepare a transformed microorganism; And
Infecting the plant with the transformed microorganism;
Wherein the target protein is expressed specifically in stem apical meristems, leaves, flowers and pods. 10. The method of claim 9,
Wherein said microorganism is Agrobacterium tumefaciens.
10. The method of claim 9,
Wherein the plant is a Arabidopsis thaliana.

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