KR101044093B1 - Gene Coding for Alanine Glyoxylate Aminotransferase and Stress-Resistant Plants Transformed by the Gene - Google Patents

Abstract

The present invention relates to an alanine glyoxylic acid aminotransferase gene and a stress resistant plant transformed with the gene, and more specifically, to alanine glyoxylate aminotransferase induced by oxidative stress. Coding green beans ( Vigna The present invention relates to a novel gene Mlti26 derived from radiata , a recombinant vector containing the gene, and a stress resistant plant transformed with the recombinant vector.

The transgenic plant according to the present invention is useful for improving crop productivity because it has resistance to oxidative stress such as low temperature, high salt, drought stress, and absic acid hormone.

Mung Bean, Mlti26, Alanine Glyoxylic Acid Aminotransferase, Oxidative Stress

Description

Gene Coding for Alanine Glyoxylate Aminotransferase and Stress-Resistant Plants Transformed by the Gene}

The present invention relates to an alanine glyoxylic acid aminotransferase gene and a stress resistant plant transformed with the gene, and more specifically, to alanine glyoxylate aminotransferase induced by oxidative stress. The present invention relates to a novel gene Mlti26 derived from mung bean ( Vigna radiata ) encoding, a recombinant vector containing the gene, and a stress resistant plant transformed with the recombinant vector.

According to the agreement, Vigna radiata ) is used to treat diseases such as single, heat, rubella, and mineral aura due to its cold nature, taste, and no poison. In addition, the pharmacology is recognized by lowering heat and swelling, lowering qi and quenching thirst. Especially, antipyretic and detoxification is excellent, and has been used as a folk medicine or cosmetics since ancient times.

On the other hand, aminotransferase, which is a kind of enzyme involved in amino acid biosynthesis and degradation, is also called transaminase, and gamma of alpha-keto acid of gamma-amino group of alpha-amino acid. It serves to catalyze the transition to the gamma-keto group.

Among the aminotransferases, alanine glyoxylate aminotransferase (EC2.6.1.2) is involved in producing glycine by transferring amino groups from amino acids such as alanine or serine to glyoxylate. Known and present in most yeast, animal and plant cells.

In addition, alanine glyoxylic acid aminotransferase catalyzes a reversible aminotransfer reaction that converts L-alanine and glyoxylic acid to pyruvate and glutamate, and increases activity by diseases such as hepatitis in animals. In plants, it has been reported to be induced by hypoxic or photorespiration. It is shown that alanine glyoxylic acid aminotransferase plays a role in removing increased L-alanine during sugar fermentation under anaerobic conditions. In the leaf tissues of C4 plants of the NAD-malic enzyme type and the phosphoenolpyruvate carboxykinase type, alanine glyoxylic acid aminotransferases were applied to the leaf meat to maintain the carbon-nitrogen balance in the C4-dicarboxylic acid pathway. It acts on the metastasis of C3 monomers between ductal sheath cells. In addition, alanine is produced in relatively high amounts in most plant roots exposed to hypoxic or anoxic conditions.

Accordingly, the present inventors have made intensive efforts to develop stress resistant plants capable of producing alanine glyoxylic acid aminotransferase under an oxidative stress environment. As a result, the novel Mlti26 gene induced by cold stress is isolated from mung bean and transformed into the gene. The present invention was completed by confirming that the plant expresses alanine glyoxylic acid aminotransferase and has resistance to oxidative stress.

The main object of the present invention is to provide a novel alanine glyoxylic acid aminotransferase gene ( Mlti26 ) induced by oxidative stress and the enzyme encoded thereby.

Another object of the present invention is to provide a transgenic plant having oxidative stress resistance to which the alanine glyoxylic acid aminotransferase gene is introduced.

In order to achieve the above object, the present invention provides an alanine glyoxylic acid aminotransferase (Mlti26) having the amino acid sequence of SEQ ID NO: 1.

The present invention also provides a gene encoding the alanine glyoxylic acid aminotransferase ( Mlti26 ), a recombinant vector comprising the gene, and a microorganism transformed with the recombinant vector.

The present invention, the Mlti26 Provided is an oxidative stress resistant plant transformed with a gene or the recombinant vector.

The present invention also comprises the steps of culturing the transformed microorganism; And it provides a method for producing alanine glyoxylic acid aminotransferase Mlti26 comprising recovering alanine glyoxylic acid aminotransferase Mlti26.

The novel alanine glyoxylic acid aminotransferase Mlti26 gene according to the present invention is induced by oxidative stress, and the transgenic plant into which the gene is introduced has oxidative stress resistance, thereby improving crop yield and quality of production. .

The present invention relates to alanine glyoxylic acid aminotransferase (Mlti26) in one aspect.

In another aspect, the present invention relates to a gene encoding the alanine glyoxylic acid aminotransferase ( Mlti26 ), a recombinant vector comprising the gene, and a microorganism transformed with the recombinant vector.

In the present invention, the gene may be characterized by having a nucleotide sequence of SEQ ID NO: 2, may be characterized by being induced by oxidative stress, the oxidative stress is low temperature, high salt, drought stress and abscis acid ( abscisic acid, ABA) may be characterized by being induced by the hormone.

In the present invention, the microorganism may be characterized in that E. coli .

In another aspect, the present invention, the Mlti26 It relates to an oxidative stress resistant plant transformed with a gene or the recombinant vector.

In the present invention, the plant is tobacco, rice, barley, wheat, soybeans, sesame, crude, sorghum, buckwheat, millet, yulmu, oats, Arabidopsis, pepper, potatoes, green beans, beans, tobacco, ginseng, rice, It may be characterized in that it is selected from the group consisting of sweet potatoes and potatoes.

In another aspect, the present invention comprises the steps of culturing the transformed microorganism; And recovering alanine glyoxylic acid aminotransferase Mlti26.

In an embodiment of the present invention, after applying cold stress to mung beans to obtain cDNA fragments induced by cold stress, the alanine glyoxylic acid aminotransferase gene is separated from the cDNA gene bank by RACE PCR, and then sequenced. Was analyzed. As a result, the isolated gene has a nucleotide sequence of 1,306bp, and confirmed that it is a novel alanine glyoxylic acid aminotransferase gene encoding 401 amino acids.

In another embodiment of the present invention, mung bean was treated at 4 ° C. low temperature, 0.1 M NaCl high salt treatment, drought stress and acetic acid hormone treatment, and then the whole RNA of the mung bean leaves was separated using HOT phenol method to perform Northern blot. It was. As a result, Mlti26 The gene was found to be a gene induced expression at low temperature, high salt, drought stress, and treatment with abscitic acid hormone.

In addition, Mlti26 After amplifying the gene by PCR, it was cloned into the pGEM T-Easy vector containing the green fluorescent protein (GFP), and then introduced into tobacco to prepare a transformant to confirm the intracellular location of the GFP-Mlti26 fusion protein. Strongly expressed in peroxysomes.

In another embodiment of the invention Mlti26 After amplifying the gene by PCR, it was inserted into pET100 / D-TOPO vector to produce a recombinant vector, and then introduced into E. coli BL21. After culturing the transformant, the supernatant was separated and subjected to electrophoresis, whereby alanine glyoxylic acid aminotransferase was expressed.

In the present invention, "vector" refers to a DNA preparation containing a DNA sequence operably linked to a suitable regulatory sequence capable of expressing DNA in a suitable host. Vectors can be plasmids, phage particles or simply potential genomic inserts. Once transformed into the appropriate host, the vector can replicate and function independently of the host genome, or in some cases can be integrated into the genome itself. Since plasmids are the most commonly used form of current vectors, "plasmid" and "vector" are sometimes used interchangeably in the context of the present invention. For the purposes of the present invention, it is preferred to use plasmid vectors. Typical plasmid vectors that can be used for this purpose include (a) a replication initiation point that allows for efficient replication to include hundreds of plasmid vectors per host cell, and (b) host cells transformed with the plasmid vector. It has a structure comprising an antibiotic resistance gene and (c) a restriction enzyme cleavage site into which foreign DNA fragments can be inserted. Although no appropriate restriction enzyme cleavage site is present, the use of synthetic oligonucleotide adapters or linkers according to conventional methods facilitates ligation of the vector and foreign DNA.

After ligation, the vector should be transformed into the appropriate host cell. Transformation is described by Sambrook, et . al ., the calcium chloride method described in section 1.82 of supra . Alternatively, electroporation (Neumann, et . Al ., EMBO J. , 1: 841, 1982) can also be used for transformation of these cells.

As the vector used for overexpression of the gene according to the present invention, an expression vector known in the art may be used.

Nucleic acids are "operably linked" when placed in a functional relationship with other nucleic acid sequences. This may be genes and regulatory sequence (s) linked in such a way as to enable gene expression when appropriate molecules (eg, transcriptional activating proteins) are bound to regulatory sequence (s). For example, the DNA for a pre-sequence or secretion leader is operably linked to the DNA for the polypeptide when expressed as a shear protein that participates in the secretion of the polypeptide; A promoter or enhancer is operably linked to a coding sequence when it affects the transcription of the sequence; Or the ribosomal binding site is operably linked to a coding sequence when it affects the transcription of the sequence; Or the ribosomal binding site is operably linked to a coding sequence when positioned to facilitate translation. In general, "operably linked" means that the linked DNA sequence is in contact, and in the case of a secretory leader, is in contact and present within the reading frame. However, enhancers do not need to touch. Linking of these sequences is performed by ligation (linking) at convenient restriction enzyme sites. If such sites do not exist, synthetic oligonucleotide adapters or linkers according to conventional methods are used.

As is well known in the art, to raise the expression level of a transfected gene in a host cell, the gene must be operably linked to transcriptional and translational expression control sequences that function in the selected expression host. Preferably, the expression control sequence and the corresponding gene will be included in one recombinant vector containing the bacterial selection marker and the replication origin. If the host cell is a eukaryotic cell, the recombinant vector must further comprise an expression marker useful in the eukaryotic expression host.

Host cells transformed with the recombinant vectors described above constitute another aspect of the present invention. As used herein, the term “transformation” means introducing DNA into a host so that the DNA is replicable as an extrachromosomal factor or by chromosomal integration.

Of course, it should be understood that not all vectors function equally well to express the DNA sequences of the present invention. Likewise not all hosts function equally for the same expression system. However, those skilled in the art can make appropriate choices among various vectors, expression control sequences and hosts without departing from the scope of the present invention without undue experimental burden. For example, in selecting a vector, the host must be considered, since the vector must be replicated in it. The number of copies of the vector, the ability to control the number of copies, and the expression of other proteins encoded by the vector, such as antibiotic markers, must also be considered.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only to illustrate the invention, it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as limited by these examples.

In particular, the following examples illustrate only tobacco as a plant transformed with the Mlti26 gene according to the present invention, but the same method of rice, barley, wheat, soybean, sesame, crude, sorghum, buckwheat, millet, yulmu, oats, Arabidopsis Pepper, Potato, Mung Bean, Bean, Tobacco, Ginseng, Paddy, It will be apparent to those skilled in the art to prepare a transformant resistant to oxidative stress by applying to sweet potatoes and potatoes.

Example  1: Mung bean-derived alanine glyoxylic acid Aminotransferase  Sequence and amino acid sequence analysis of genes

Mung beans grown for more than 3 weeks in growth were treated with 0, 3, 6, 12, 24 hours in a 4 ° C low temperature incubator to induce low temperature stress, and then obtained mung bean leaves to extract poly (A +) mRNA, followed by PCR- Select cDNA subtraction (Clontech) was used to obtain cDNA fragments expressed by cold stress.

The obtained cDNA fragment was cloned into the pCR2.1 vector (Invitrogen), which is a subcloning vector, and plasmid DNA was isolated using a plasmid purification kit (NucleoGen). Subsequently, sequencing of the isolated plasmid DNA was performed by requesting Macrogen, and comparative analysis of sequencing was performed by DNASIS and BLAST programs provided by NCBI, GenBank, EMBL, and SwissProt databases. Based on the analyzed sequencing results, the entire nucleotide sequence was obtained using RACE-PCR.

As a result, it was confirmed that the cDNA of the alanine glyoxylate aminotransferase gene induced by cold stress is composed of a total of 1,306 bp sequences (SEQ ID NO: 2). 401 amino acids were deduced (SEQ ID NO: 1). The nucleotide sequence was found to be a novel gene induced by cold stress, which was named Mlti26 .

Example  2: Mung bean genomic Mlti26 gene amount  black

Genomic DNA was extracted from the leaf tissue of mung beans, treated with restriction enzyme Nco I or BstX I, followed by electrophoresis on a 0.8% agarose gel, and then the gel was denaturation solution (0.5M NaOH, 1.5M NaCl). ). Subsequently, it was immersed in neutralization solution (0.5 M Tris-Cl, 1.5M NaCl, pH 7.5), and then transferred to nylon membrane (AP Biotech) with 10 × SSC solution (1.5M NaCl, 0.15M sodium citrate). Southern blot was performed using alanine glyoxylic acid aminotransferase gene fragment (1,306 bp cDNA) labeled with radioisotope [a- ³² P] dCTP (FIG. 1).

As a result, one band was detected when the restriction enzyme EcoR I was treated, and two bands were detected when the restriction enzyme BstX I was treated (FIG. 1). Thus, alanine glyoxylic acid aminotransferase cDNA has no site that recognizes restriction enzyme EcoR I, one site that recognizes restriction enzyme BstX I exists, and there is a single copy of Mlti26 gene on the mung bean genome. Could know.

Example  3: Mlti26  Expression by Gene Oxidative Stress

The expression of the Mlti26 gene isolated in Example 1 at low temperature, high salt, drought stress and Abscisic acid (ABA) hormone treatment was confirmed.

That is, mung beans grown for three weeks or more were grown at a low temperature of 4 ° C. by the method described in Example 1 above, or 0.1M NaCl was treated for 6, 12, and 24 hours. On the other hand, in order to induce drought stress, the soil of mung bean root was removed and treated for 6, 12, 24 hours in a 28 ℃ incubator. In addition, 1 mM Apsy acid hormone (Sigma, USA) was sprayed on mung bean plants and treated for 6, 12, 24 hours. Subsequently, the total RNA of the mung bean leaves was isolated using the HOT phenol method (Verwoerd et. al ., Nucl . Acid . Res ., 7: 2362, 1989).

The total RNA isolated above was electrophoresed on a 1.0% formaldehyde gel, and the gel was subjected to a nylon membrane (Schleicher & Schuell) with 20 × SSC solution (3M NaCl, 0.3M sodium citrate). Transfer was performed, and Northern blot was performed using a fragment (1,306 bp cDNA) of the alanine glyoxylic acid aminotransferase gene labeled with the radioisotope [a- ³² P] dCTP.

As a result, it was confirmed that the transcript was strongly expressed from 3 hours after the low temperature treatment, and after 12 hours, the amount of RNA expression was slightly decreased, and then the most transcript was expressed at 24 hours (FIG. 2A).

When NaCl was treated, significant transcripts were induced after 6 hours, but after that transcript expression decreased significantly (FIG. 2B).

The drought stress gradually increased the amount of transcript, and a significant amount of transcript was expressed up to 24 hours (FIG. 2B).

In addition, in the case of the treatment with abscitic acid hormone, the transcript was strongly expressed after 6 hours, and the expression level of the transcript decreased as time passed (Fig. 2B).

Therefore, it was found that the Mlti26 gene is a gene induced expression at low temperature, high salt, drought stress, and acetic acid hormone treatment.

Example  4: using plant expression vectors GFP  Intracellular Positioning of Recombinant Fusion Proteins

Mlti26 isolated from above The cDNA of the gene was PCR using 5SacI-Mlti26 primer (SEQ ID NO: 3) and 3SacI-Mlti26 primer (SEQ ID NO: 4).

SEQ ID NO: 5'-GAGCTCATGGATTATTTCAATGCAC

SEQ ID NO: 5'-GAGCTCTCAAATCCTGGAAGGG

The PCR fragment amplified above was cloned into a pGEM T-Easy vector (Promega) containing a green fluorescence protein (GFP) as a fluorescent marker in a plant expression binary vector and confirmed by DNA sequencing. . The restriction enzyme Sac I-inserted Mlti26 DNA fragment was inserted into the restriction enzyme Sac I site of the pGEM T-Easy vector, thereby fusion with the C-terminal portion of GFP (FIG. 3A). The recombinant vector was introduced into tobacco to induce the expression of the GFP-Mlti26 fusion protein.

TargetP ( http://www.cbs.dtu.dk/services/Target P /), an internet protein location analysis program, and PSORT II ( http://psort.ims.u-tokyo.ac.jp/form2.html ) As a result of the analysis, the mungbean Mlti26 protein was predicted to be targeted as a peroxisome, and the intracellular location of the GFP-Mlti26 fusion protein in the tobacco protoplasts into which the recombinant vector was introduced was determined by fluorescence microscopy. It was found to be strongly expressed in oxysomes (FIG. 3B).

Example  5: Confirmation of Recombinant Fusion Protein Expression Using Escherichia Coli Expression Vectors

CDNA of the Mlti26 gene isolated above was subjected to PCR using CACC-Mlti26 primer (SEQ ID NO: 5) and 3-Mlti26 primer (SEQ ID NO: 6).

SEQ ID NO: 5'-CACCATGGATTATTTCAATGCAC

SEQ ID NO: 5'-TCAAGTGTTCTGTAAGTATGCACTGG

The PCR fragment was inserted into the sites of restriction enzymes BamH I and EcoR I of pET100 / D-TOPO (Invitrogen), His-tag fusion vectors, of the E. coli expression vector, whereby the T7 promoter and isopropyl-thio-β-D-gal- Expression of recombinant fusion protein (pET-Mlti26, 47 kDa) was induced under the control of isopropyl-thio-β-D-gal-actopyranoside (IPTG) (FIG. 4A).

In order to express the Mlti26 gene in Escherichia coli, the recombinant vector prepared above was inserted into Escherichia coli BL21 with a temperature shock (42 ° C), and then in a LB (Luria-Bertani) liquid medium containing 75 µg / ml Amuchcillin (Duchafa). After incubation for 1 hour, the plate was plated in LB-Amp solid medium and incubated for 16 hours at 37 ° C.

The cultured bacteria were incubated in 50 ml LB liquid medium containing ampicillin to OD 0.6 (1.0 × 10 ⁶ cells), incubated for 3 hours at 28 ° C. with 0.1 mM IPTG, or 14 ° C. with 0.1 mM IPTG. Incubated for 20 hours at. After the incubation, cells were obtained by centrifugation at 3,000 rpm for 5 minutes, and then 1.7 ml of 50 mM Tris-HCl (pH 8.0) buffer was added thereto. The cells were disrupted using a cell crusher and centrifuged at 5,000 rpm for 5 minutes.

After the centrifugation, the supernatant was separated and again centrifuged at 12,000 rpm for 10 minutes, and water-soluble and non-water-soluble proteins were separated to obtain 12% SDS-protein electrophoresis (Laemmli, Nature , 227: 680, 1970). Recombinant fusion alanine glyoxylic acid aminotransferase protein expression was analyzed (FIG. 4B). As a result, more fusion protein was expressed when cultured at 14 ° C. than at 28 ° C. (FIG. 4B).

In addition, in order to confirm that His-Mlti26 protein is actually expressed, Western blot was performed using His-tag monoclonal antibody (Invitrogen), and both His-Mlti26 and 28C were cultured at 14 ° C. It was confirmed that the fusion protein is strongly expressed at the 47kDa position (FIG. 4C).

As a result, it was found that His-Mlti26 protein could be efficiently expressed in E. coli using the His-tag fusion protein expression system.

As described above in detail specific parts of the present invention, it is apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

1 is a Southern blot to determine the number of alanine glyoxylic acid aminotransferase gene ( Mlti26 ) isolated from mung beans in the mung bean genome.

Figure 2 is to perform the Northern blot to isolate the total RNA from low-temperature, high salt, drought stress and ABA hormone-treated mung bean to confirm the expression of the Mlti26 gene.

Figure 3 shows the expression of GFP recombinant protein of the protein encoded by the Mlti26 gene in tobacco plasma cells to analyze the location (A: showing the fusion form of the GFP-Mlti26 fusion protein; B: the recombinant vector is introduced Intracellular locations of GFP (left) and GFP-Mlti26 fusion proteins (right) in tobacco protoplasts confirmed by fluorescence microscopy.

Figure 4 is a picture of a large amount of recombinant protein containing Mlti26 gene in E. coli and analyzed by protein electrophoresis (SDS-PAGE) and Western blot.

<110> Technology Licensing Center Dong-A University <120> GENE CODING FOR ALANINE GLYOXYLATE AMINOTRANSFERASE AND          STRESS-RESISTANT PLANTS TRANSFORMED BY THE GENE <160> 6 <170> KopatentIn 1.71 <210> 1 <211> 401 <212> PRT <213> Mlti26 <400> 1 Met Asp Tyr Phe Asn Ala Pro Gly Arg Asn His Leu Phe Val Pro Gly   1 5 10 15 Pro Val Asn Ile Pro Asp Gln Ile Ile Arg Ala Met Asn Arg Asn Asn              20 25 30 Glu Asp Tyr Arg Ser Pro Ala Ile Pro Ala Met Thr Lys Thr Leu Leu          35 40 45 Glu Asp Val Lys Lys Ile Phe Lys Thr Thr Thr Gly Thr Pro Phe Leu      50 55 60 Ile Pro Thr Thr Gly Thr Gly Ala Trp Glu Ser Ala Leu Thr Asn Thr  65 70 75 80 Leu Ser Pro Gly Asp Arg Ile Val Ser Phe Leu Ile Gly Gln Phe Ser                  85 90 95 Leu Leu Trp Ile Asp Gln Gln Gln Arg Leu Lys Phe Asn Val Asp Val             100 105 110 Val Glu Ser Glu Trp Gly His Gly Ala Lys Leu Asp Val Leu Glu Ser         115 120 125 Lys Ile Ala Ser Asp Thr Ser His Thr Ile Lys Ala Ile Cys Ile Val     130 135 140 His Asn Glu Thr Ala Thr Gly Val Thr Asn Asp Leu Ala Lys Val Arg 145 150 155 160 Gln Ile Leu Asp Ser Tyr Gln His Pro Ala Leu Leu Ile Val Asp Gly                 165 170 175 Val Ser Ser Ile Cys Ala Leu Asp Phe Arg Met Asp Glu Trp Gly Val             180 185 190 Asp Val Ala Ile Thr Gly Ser Gln Lys Ala Leu Ser Leu Pro Thr Gly         195 200 205 Ile Gly Ile Val Val Ala Gly Pro Lys Ala Ile Glu Ala Ser Lys His     210 215 220 Ala Lys Ser Leu Arg Val Phe Phe Asp Trp Lys Asp Tyr Leu Lys Phe 225 230 235 240 Tyr Gln Leu Gly Thr Tyr Trp Pro Tyr Thr Pro Ser Ile His Leu Leu                 245 250 255 Tyr Gly Leu Arg Ala Ala Leu Asp Leu Ile Phe Glu Glu Gly Leu Glu             260 265 270 Asn Val Ile Ala Arg His Ser Arg Leu Gly Lys Ala Thr Arg Leu Ala         275 280 285 Val Glu Ala Trp Gly Leu Lys Asn Cys Thr Gln Lys Glu Glu Trp Tyr     290 295 300 Ser Asp Thr Val Thr Ala Val Leu Val Pro Ala Tyr Ile Asp Ser Thr 305 310 315 320 Glu Ile Val Arg Arg Ala Trp Lys Arg Tyr Asn Leu Ser Leu Gly Leu                 325 330 335 Gly Leu Asn Lys Val Ala Gly Lys Val Phe Arg Ile Gly His Leu Gly             340 345 350 His Leu Asn Glu Leu Gln Leu Leu Gly Cys Leu Ala Gly Val Glu Met         355 360 365 Ile Leu Lys Asp Val Gly Tyr Pro Val Lys Leu Gly Ser Gly Val Ala     370 375 380 Ala Ala Ser Ala Tyr Leu Gln Asn Thr Ile Pro Met Ile Pro Ser Arg 385 390 395 400 Ile     <210> 2 <211> 1206 <212> DNA <213> Mlti26 <400> 2 atggattatt tcaatgcacc aggaagaaac catctctttg ttccagggcc ggttaacatt 60 ccggaccaga tcatccgggc catgaacaga aacaatgagg actaccgttc tccagcaatt 120 ccagctatga caaaaacttt gcttgaggat gtcaagaaga ttttcaagac cactactgga 180 accccatttc tcatccctac aactggtact ggtgcttggg agagtgctct cacaaacaca 240 ctgtctcctg gggatcgaat tgtatcgttc ctgattggcc aattcagctt gctttggatt 300 gatcagcagc aacgcctgaa gttcaatgtt gatgttgttg agagtgaatg gggccatggt 360 gctaagcttg atgttctgga atcaaagatt gcttcagata cttcacacac tataaaggca 420 atttgcattg ttcacaatga gactgcaact ggggtcacca atgacttggc caaagtgaga 480 caaattcttg attcctacca gcatccagcc ctccttattg ttgatggagt gtcttccatt 540 tgtgctcttg atttccgcat ggatgaatgg ggagttgatg tggcaataac tggctcccag 600 aaggcccttt cccttcccac tgggataggt attgtggttg caggccctaa agctattgag 660 gcctcaaaac atgctaaatc acttagagtt ttctttgact ggaaagacta cctcaaattc 720 taccagctag gaacctattg gccatacact ccttccatac atttgctgta tggtctgaga 780 gctgctcttg atctgatttt tgaggaagga ctcgaaaatg tgatcgcaag gcacagtcgt 840 ttaggcaaag caaccagact tgctgtagag gcatggggtt tgaagaattg cacccaaaag 900 gaagagtggt acagtgacac tgtgactgct gttcttgttc ctgcttacat tgatagtact 960 gaaatagtta ggagggcatg gaagagatac aatttgagct taggtcttgg actgaacaaa 1020 gttgctggga aggttttcag aattggacat cttggccact tgaatgagtt gcaactgttg 1080 ggatgtctag ctggtgtaga gatgatactc aaagatgtgg gttatcctgt aaagcttgga 1140 agtggagttg ctgctgccag tgcatactta cagaacacta ttcctatgat cccttccagg 1200 atttga 1206 <210> 3 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 gagctcatgg attatttcaa tgcac 25 <210> 4 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 gagctctcaa atcctggaag gg 22 <210> 5 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 caccatggat tatttcaatg cac 23 <210> 6 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 6 tcaagtgttc tgtaagtatg cactgg 26  

Claims (12)

delete delete delete delete delete delete Alanine glyoxylic acid aminotransferase having the amino acid sequence of SEQ ID NO: 1 and having properties induced by one or more oxidative stresses selected from the group consisting of low temperature, high salt, drought stress and abscisic acid (ABA) hormones An oxidative stress resistant plant transformed with a gene encoding (Mlti26). The method of claim 7, wherein the plant is tobacco, rice, barley, wheat, soybeans, sesame, crude, sorghum, buckwheat, millet, pearl barley, oats, Arabidopsis, red pepper, potatoes, green beans, soybeans, tobacco, ginseng, rice, A transformed oxidative stress resistant plant, characterized in that it is selected from the group consisting of sweet potatoes and potatoes. delete delete delete delete

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