KR101639102B1 - Kinase gene resistant to a bacterial blight, and the transgenic plant using the same - Google Patents

Abstract

The present invention relates to a plant transformant transformed such that LOC_Os10g35040, an inherently inactive gene, is activated, a recombinant vector containing an expression inducing sequence for expressing the gene, and a method for producing the transformant. Further, the present invention relates to a method for promoting rice disease resistance, which comprises activating the LOC_Os10g35040 gene. The present invention relates to a transgenic rice having a strong resistance to blight of blight by inserting an expression inducing sequence into an expression regulatory region of LOC_Os10g35040, a resistance gene against blight of blight that was inherently present in rice but was not active. Transgenic rice can be widely used because it can reduce the use of pesticides, improve productivity, and provide safe agricultural products through disease resistance.

Description

[0001] The present invention relates to a gene for resistance to blight of blight of leaf blight and a transgenic plant using the same,

The present invention relates to a plant transformant transformed such that LOC_Os10g35040, an inherently inactive gene, is activated, a recombinant vector containing an expression inducing sequence for expressing the gene, and a method for producing the transformant. Further, the present invention relates to a method for promoting rice disease resistance, which comprises activating the LOC_Os10g35040 gene.

Rice blight caused by Xanthomonas oryzae pv. It is a bacterial tuberculosis caused by oryzae , and its onset occurs from early July to harvest season, mainly from early July to mid-August. Bacterial wintering is possible in underground stalks, roots, or diseased rice straw and rice stubble, which are intermediate host plants. Bacteria spread along the water by flagella and invade into the water holes, air holes and cut roots of the leaves. Disease occurs mainly in the area of frequent occurrence of the disease every year, but it also occurs in areas not in the area of the pelvic area.

Usually it appears before and after the heading period, but the occurrence of the rash or the occurrence of the rash occurs at the beginning of the reply. The first occurrence of the disease in Korea was first reported in 1930 in Haenam, Jeollanam-do. Since 1960, a limited occurrence has been observed in some parts of the southern part of the country. The cultivation of Gyeongnam cultivar showed a nationwide occurrence. 23, and it is considered to be one of the three major diseases of rice, showing not only the onset of leaf in later stage during the cultivation period but also the symptom of gold mold in which the whole body occurs until after the transition.

The rice blotting bacteria are classified as Gram negative bacteria and are widely distributed throughout the world because of bacterial fungi causing bacterial fake disease of rice. Although they cause great damage to rice, pathogens propagate in the ducts of rice, And there is no specific fungicide to date.

In addition, the conventional method of imparting resistance to crops is a technique in which plants are given new traits through the introduction of foreign genes.

Meanwhile, Dongjinbyeong was developed in 1975 as a cross breed between gold southern wind, Nakdong rice and Sadominori in 1975 with the goal of raising the disease, quality and diversity of new varieties. It is resistant to streak blight in case of disease resistance, , And it is weak against blight of white blotch and other pests. Therefore, it is required to study resistance to blight of white blossom and blast disease in Dongjinbyeong, and to cultivate improved varieties. In addition, Hwangyoung Rye is a variety cultivated by crossing Jurakes 830 with YR4811Acp 8, which is strong against blast and streak blight, in order to improve useful qualities such as good quality, tolerance to disease and multiparity. It is a varieties resistant to blast, However, it is necessary to study on the resistance resistance enhancement continuously when considering the rapid resistance of outbreaks or germs.

Accordingly, the present inventors have made intensive researches to identify a resistance gene against the blight of rice blight that has not yet been overcome. As a result, the inventors of the present invention found that a gene encoding a phosphorylated enzyme gene (LOC_Os10g35040 ), And confirmed that the transgenic plant overexpressing the transformant exhibited resistance to blight of blight, thus completing the present invention.

One object of the present invention is to provide a plant transformant which is transformed to activate the endogenous inactive gene LOC_OslOg35040.

Another object of the present invention is to provide a recombinant vector for enhancing the disease resistance of plants, which comprises an expression inducing sequence that expresses an LOC_OslOg35040, an inherent inactive gene.

It is still another object of the present invention to provide a method for producing a transformed plant having enhanced disease resistance, which comprises the step of obtaining rice cells transformed with the recombinant vector for promoting disease resistance.

Yet another object of the present invention is to provide a method for promoting rice disease resistance comprising activating the LOC_Os10g35040 gene.

In one aspect of the present invention, the present invention provides a plant transformant transformed so that LOC_Os10g35040, an inherent inactive gene, is activated.

In the present invention, a transgenic rice having resistance to blight of blight was prepared by inserting an expression inducing sequence into the expression control region of LOC_OslOg35040, a gene that was inherently present in rice but was not active.

The term "intrinsic" in the present invention means that a living organism has a natural state. In the present invention, when a plant, preferably a genus of a rice (Oriza sp.) Chromosome, use.

In the present invention, the term "inherent inactive gene" means DNA, that is, it exists as a gene but can not be transcribed into RNA or can not be translated into protein, This means that the gene inherently possessed by rice does not express its function due to the transcription or non-transcriptional nature, and in particular, the LOC_Os10g35040 gene does not express its function.

In the present invention, the term "LOC_Os10g35040" is a gene present in the 10th chromosome of Oryza sativa, and its expression product is a precursor of protein kinase receptor and is a kind of nRD phosphorylase.

In a specific embodiment of the present invention, the LOC_Os10g35040 gene was found to be inactivated in the native rice (FIG. 3), and the expression of the LOC_Os10g35040 gene was specifically induced in the transformant (FIG. 4), and it was confirmed that the plant was resistant to blight of blight (FIG. 5).

In the present invention, the inherently inactive gene may preferably be a LOC_Os10g35040 gene, more preferably a gene consisting of a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 1, and most preferably, The nucleotide sequence of SEQ ID NO.

In the present invention, the term "activating" means enhancing the activity of a specific gene, and includes 1) introducing at least one copy of the corresponding gene, 2) inserting a sequence inducing expression in an expression- By enrichment in a manner that transforms the activity into an enhanced form, or 4) by a combination thereof. In the present invention, it is preferable to insert a sequence or a promoter which induces expression in an expression control region of an endogenous gene, more preferably a sequence including an expression induction sequence of SEQ ID NO: 9 have. Most preferably, the sequence described in SEQ ID NO: 2 can be inserted.

The term "expression inducing sequence" as used herein includes, without limitation, a sequence which functions to promote expression when it is present in a gene expression regulatory region and includes a transcription factor binding site, an enhancer, a co- binding region). In the present invention, it is preferable that the sequence inserted into the expression control region of the endogenous inactive gene includes the expression induction sequence of SEQ ID NO: 9. More preferably, it may be composed of the nucleotide sequence shown in SEQ ID NO: 2.

In a specific embodiment of the present invention, it was confirmed that the gene of LOC_Os10g35040 gene was expressed by inserting an expression inducing sequence (SEQ ID NO: 2) into the expression control region of the LOC_Os10g35040 gene (FIG. 3).

The term "transformed" in the present invention means all the activities in which a specific trait occurs, disappears or is controlled through mutation such as insertion, deletion or substitution on the DNA base sequence. In particular, in the present invention, Which means that the genetic properties of In the present invention, the term "transformant" refers to a transgenic plant produced by transformation, and includes genetic recombinants produced by mutation of a specific gene or activation of an activity using genetic recombination technology.

In the present invention, the term "plant transformant" includes all plants which can be subjected to transformation, and in particular, all plants in which the LOC_Os10g35040 gene or its corresponding gene is inactivated are all included without limitation. Preferably, the plant transformant may be rice (Oryza sp.), And more preferably, it may be Dong-Jin-jin or a chemical fertilizer.

In the present invention, the plant transformant may have resistance to blight of blight.

In one specific embodiment of the present invention, the expression inducing sequence of SEQ ID NO: 2 is inserted into the expression-regulating region of LOC_Os10g35040, which is an inherent inactivation gene of the present invention, to transform the gene expressing LOC_Os10g35040 , And named it nRDK-08.

In another aspect, the present invention provides a recombinant vector for promoting disease resistance of a plant, which comprises an expression inducing sequence that expresses an intrinsic inactive gene, LOC_Os10g35040.

The intrinsic inactive gene, LOC_OslOg35040 and the expression inducing sequence are as described above.

In the present invention, the term "disease resistance" means that the onset of disease does not occur well, or that the disease progression rate is significantly slower or recoverable. In the present invention, disease resistance may mean resistance to blight of blight.

In the present invention, the term "recombinant vector" means a vector that has undergone a gene recombination process in which a specific sequence is mutated, inserted or deleted, preferably a vector into which an expression inducing sequence is inserted, Means a recombinant vector capable of inserting a sequence into the expression control region of LOC_Os10g35040.

In one embodiment of the present invention, a recombinant vector containing T-DNA was prepared, introduced into rice cells through Agrobacterium, and the T-DNA sequence was inserted into the expression control region of LOC_Os10g35040 to generate LOC_Os10g35040 Expression of the gene was induced (FIG. 3).

In another embodiment, the present invention provides a recombinant vector comprising the steps of: introducing a recombinant vector for transformation into an Agrobacterium containing an expression inducing sequence that expresses an intrinsic inactive gene LOC_Os10g35040; A step of infecting the rice leaves with the Agrobacterium, and a step of callusing the infected rice cells to obtain transformed rice cells.

The term "Agrobacterium" in the present invention refers to a gram-positive bacterium, which is infected with plant cells through scars of the roots or stems of plants, and inserts the gene of interest into the genome of the plant cell to transform the plant cell Is a kind of carcinoma bacteria that causes cancer. In the present invention, the Agrobacterium may be Agrobacterium tumefaciens.

The term "callus " in the present invention means an undifferentiated amorphous cell mass produced when a plant is injured, that is, undifferentiated cell. Particularly, in the present invention, Agrobacterium When the transforming carcinoma bacterium is infected, it means that the infected cells are undifferentiated and lump on the wound site. That is, in the present invention, transformed undifferentiated cells can be obtained using only the cells transformed by infection with carcinoma bacteria to produce a mass of carcinoma, which is produced as a callus.

The method for producing a transformant having enhanced disease resistance may further include the step of differentiating the obtained callus into root and adult. Since the callus is only a cell mass in an undifferentiated state as described above, it may be necessary to induce differentiation into roots and adults in order to obtain it as a whole seed.

In one specific embodiment of the present invention, the resulting callus was induced to differentiate into root and adult by culturing in root-induction medium.

In another aspect, the present invention provides a method for enhancing rice disease resistance comprising activating the LOC_Os10g35040 gene.

The LOC_Os10g35040 gene, activation and disease resistance of the present invention are as described above.

The present invention relates to a transgenic rice having a strong resistance to blight of blight by inserting an expression inducing sequence into an expression regulatory region of LOC_Os10g35040, a resistance gene against blight of blight that was inherently present in rice but was not active. Transgenic rice can be widely used because it can reduce the use of pesticides, improve productivity, and provide safe agricultural products through disease resistance.

Fig. 1 is an outline of the structure of LOC_Os10g35040 selected as a gene related to resistance to blight of blight. Fig. 1 (A) shows the structure of the gene phase. Fig. 1 (B) shows the structure of the gene-expressing protein. The LRR represented by this indicates leucine-rich repeats, TM means a transmembrane domain, and PK means a protein kinase.
Fig. 2 is a structural diagram showing a part of the genomic DNA of nRDK-08 transgenic rice in which a gene expression induction sequence including T-DNA is inserted in a region regulating LOC_Os10g35040 expression.
FIG. 3 is a diagram showing RT-PCR results confirming the expression of LOC_Os10g35040 in nRDK-08 transgenic rice.
FIG. 4 is a graph showing that expression of both genes of LOC_Os10g35040 in nRDK-08 transgenic rice was not changed by genetic modification but only LOC_Os10g35040 gene was specifically induced.
FIG. 5 is a graph showing the results of an experiment for resistance to blight of blight of nRDK-08 transgenic rice, and FIG. 5A shows the results of inoculation of blunt-ended blight of naturally occurring rice and nRDK-08 transgenic rice with scissors And FIG. 5B is a graph showing the results of measuring the lengths of rice leaves destroyed by infection thereof.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to illustrate the present invention, but the scope of the present invention is not limited thereto.

Example 1. Screening of Transgenic Target Gene through Bioinformatics

We identified 448 non-arginine-aspartate (nRD) phosphorylase enzymes, which are known to be closely related to the disease resistance of plants possessed by the genome of Dongjin / Of these, 61 genes with high probability of resistance to blight of blight were selected using the bioinformatics tool (Web site: http://www.ricearray.org, Program: Mev- Multi Experimet Viewer) (* p <1.8 ). In the process of isolating genes having disease resistance by preparing transformants for selected genes, LOC_Os10g35040 of the present invention was selected as a gene resistant to blight of blight.

In this process, only 5 genes from 448 nRD phosphorylases were novel genes related to disease resistance. Among the inherent disease resistance-related candidate nRD phosphorylase genes of the rice, only 1.1% of the genes actually associated with disease resistance were found.

Example 2. Preparation of transgenic rice with expression derivatives

2-1. Production of recombinant vector for rice transformation

A vector was constructed to insert an expression inducing sequence into the nucleotide sequence of the expression control region of the target gene.

First, the T-DNA for inserting the genome, the N-GUS for identifying the left gene sequence at the insertion site, and the recombinant vector containing the expression induction sequence of the gene of SEQ ID NO: 2 were obtained from Tobacco Leaf Mosaic Virus 35S promoter DNA, and a vector having L-0.5 / 1.5 to confirm the dhfmsWHr gene sequence at the insertion site. The primers for identification and amplification of the target gene from the inserted genome are as follows.

Enhancer-F (forward, SEQ ID NO: 3)

5'-ccggtaccATGGCGGCGGTGGTGGAGGG-3 '

Enhancer-R (reverse, SEQ ID NO: 4)

5'-ccggatccTTAGCTTAGAGATGAGACGAA-3 '

The PCR amplification conditions were 1 cycle at 94 ° C for 3 minutes, 1 cycle at 94 ° C for 1 minute, 1 minute at 58 ° C and 2 minutes at 72 ° C for thermal denaturation of DNA for the first time, Respectively.

The amplified PCR product was electrophoresed with 1.0% agarose gel, stained with EtBr (ethidium bromide) solution, and the band corresponding to the base size was cut under UV lamp to obtain agarose gel purified &Lt; / RTI &gt;

The extracted PCR product was digested with the restriction enzyme BamHI / KpnI. Further, the recombinant vector pGA2715 vector was digested with restriction enzyme BamHI / KpnI, and the digested vector was purified by the above agarose gel purification method. A DNA fragment from the PCR product was inserted into the cleaved recombinant vector to prepare a recombinant vector containing the derived expression vector.

2-2. Transformant production

The prepared vector was introduced into Agrobacterium tumefaciens using electroporation. The seedlings grown in the aseptic condition in the culture room were transformed by leaf disc method. Specifically, the rice leaves were cut into approximately 5 x 5 mm size and placed in an MSO liquid medium (4.3 g of MS salt, 30 g / L of sucrose, pH 5.6 to 5.8) containing 200 μl of Agrobacterium, The cells were cultured for 3 days and the cultured leaf slices were seeded in callus-shoot induction medium (100 μg NAA, 1 mg BA, 500 μg cell supernatant, 1 mg phospinothricin, 0.7% phytoplankton, Phyto agar, pH 5.6) and transferred to fresh medium. Induced germination was transferred to root-induction medium (250 μg of cell suspension, 10 mg phosphinotricin, 0.7% phyto-agar, pH 5.6) in MSO medium if it grew to about 1.5 to 2 cm.

The gene transgenic rice in which the expression derivative was inserted was named nRDK-08, and its gene structure is as shown in Fig. Specifically, a transformant having a dielectric structure as shown in FIG. 2 was prepared by inserting a T-DNA sequence into an upstream region predicted to be an expression-regulating region of LOC_Os10g35040 present in the 10th genome of Dong Jinbin.

2-3. Identification of rice embryo insertion position of recombinant vector

The insertion site of the vector for inducing the expression of the target gene in the genome of the transformant was confirmed by using a primer specific to the end of the inserted vector.

The primer sequences used here are as follows.

pGA2715L1.5S-F (SEQ ID NO: 5)

5'-AGTACATTAAAAACGTCCGC-3 '

As a result of confirming using the above primer, it was confirmed that the insertion position of the recombination vector was a site for regulating the expression of the target gene.

Example 3. Verification of Genetic Activity of Transgenic Rice

The gene activity of LOC_Os10g35040 was analyzed by RT-PCR (reverse-transcriptase-PCR) in transgenic rice (nRDK-08) transfected with expression derivatives in the expression control region of LOC_Os10g35040 gene, Respectively.

Specifically, total RNA was extracted from 0.1 g of the leaves of transformed rice (nRDK-08). 5'-GTCGCCTCCCCGTCTC-3 ', SEQ ID NO: 7) and a reverse transcriptase (reverse transcriptase) Reverse transcriptase PCR was performed using DNA polymerase.

As can be seen from FIG. 3, LOC_Os10g35040 was not expressed in the wild-type Hwangyong, but LOC_Os10g35040 was expressed in the transgenic rice of the present invention.

On the other hand, in order to confirm that LOC_Os10g35040-specific activity was induced, expression of a neighboring gene existing in both LOC_Os10g35040 and LOC_Os10g35040 was confirmed by reverse transcriptase PCR (FIG. 4).

As shown in FIG. 4, the expression of the LOC_Os10g35040 gene was induced in the transformant nRDK-08 in which the expression of LOC_Os10g35040 was induced by the insertion of the expression inducing sequence, while the neighboring genes existing on both sides of the gene were expressed . As a result, it was confirmed that expression of the LOC_Os10g35040 gene was induced only by the insertion of the expression inducing sequence and did not affect other genes.

Example 4: Resistance test of transformed rice (nRDK-08) against rice blight blight of rice

In order to confirm whether the transformed rice produced in Example 2 showed resistance to blight of blight, nRDK-08, a transgenic rice with expression derivatives inserted into the LOC_Os10g35040 expression control region, was used as a control, and natural rhodopsin and other nRD phosphorylase Xanthomonas oryzae pv. Strain, which induces blight of blight, was grown for 6 weeks . oryzae (strain: PXO99Az) scissors were inoculated two weeks after inoculation (Fig. 5). Lesion lengths were measured by infected and destroyed lesions from inoculated sites and compared with other nRD phosphorylase transformants that were found to be unrelated to wild-type and other disease resistance.

As shown in Fig. 5B, the nRDK-08 transformant had only half of the average destructed wound length of natural or other nRD phosphorylase transformants (naturally occurring vs. nRDK-08: 13.5 vs. nRDK-08). 7.1), and it was confirmed that it has a strong resistance to blight of blight.

From these results, it was confirmed that the transformed rice nRDK-08 of the present invention can be resistant to blight of blight by activating the intrinsic gene LOC_Os10g35040 which was not activated.

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

<110> University-Industry Cooperation Group of Kyung Hee University <120> Kinase gene resistant to a bacterial blight, and the transgenic          plant using the same <130> PA120851KR <160> 9 <170> Kopatentin 2.0 <210> 1 <211> 642 <212> PRT <213> Oryza sativa LOC_Os10g35040 <400> 1 Met Ala Ala Val Val Glu Gly Val Gly Trp Pro Glu Pro Glu Leu Glu   1 5 10 15 Ala Leu Arg Asp Ala Leu Arg Asp Ala Leu              20 25 30 Arg Ser Gly Arg Asp Leu His Ser Asn Trp Thr Gly Pro Pro Cys His          35 40 45 Gly Gly Arg Ser Ser Arg Tyr Gly Val Ala Cys Asp Gly Asp Gly Arg      50 55 60 Val Val Gly Val Gln Leu Asp Gly Ala Gln Leu Thr Gly Ala Leu Pro  65 70 75 80 Ala Gly Ala Leu Ala Gly Val Ala Arg Leu Glu Thr Leu Ser Leu Arg                  85 90 95 Asp Asn Ala Ile His Gly Ala Leu Pro Arg Leu Asp Ala Leu Ala Arg             100 105 110 Leu Arg Val Val Asp Leu Ser Ser Asn Arg Phe Ser Gly Pro Ile Pro         115 120 125 Arg Gly Tyr Ala Ala Leu Gly Glu Leu Thr Arg Leu Glu Leu Gln     130 135 140 Asp Asn Leu Ile Asn Gly Thr Leu Pro Ala Phe Glu Gln Asp Gly Leu 145 150 155 160 Ala Val Phe Asn Val Ser Tyr Asn Phe Leu Gln Gly Glu Val Pro Asp                 165 170 175 Thr Arg Ala Leu Arg Arg Phe Pro Ala Thr Ala Phe Ala His Asn Leu             180 185 190 Arg Leu Cys Gly Glu Val Val Arg Thr Glu Cys Arg Arg Glu Gly Ser         195 200 205 Pro Phe Asp Ala Ala Pro Ala Gly Gly Gly Gly Ser Gly Ser Asp Gly     210 215 220 Gly Asp Arg Val Phe Gly Ala Arg Asp Ala Ala Ala Pro Pro Ala Arg 225 230 235 240 Trp Arg Lys Pro Ile Arg Phe Arg Ile Ala Arg Trp Ser Val Val Val                 245 250 255 Ile Ala Leu Ile Ala Leu Ile Phe             260 265 270 Leu His His Ser Lys Lys Ser Arg Val Asp Lys Ala Ala Glu Gln Ala         275 280 285 Gly Lys Lys Val Ser Ser Gly Ser Gly Asn Gly Ser Ser Ser Thr Thr     290 295 300 Glu Ser Gly Lys Gly Ala Ala Asp Gln Leu Gln Phe Phe Arg Pro Glu 305 310 315 320 Lys Ala Thr Phe Ser Leu Asp Glu Leu Phe Arg Ser Thr Ala Glu Met                 325 330 335 Leu Gly Lys Gly Arg Leu Gly Ile Thr Tyr Arg Val Ala Leu His Ala             340 345 350 Gly Gly Gly Gly Gly Gly Gly Gly Gly Pro Val Val Val Val Lys         355 360 365 Arg Leu Arg Asn Met Gly His Val Pro Arg Lys Asp Phe Ala His Thr     370 375 380 Met Gln Leu Leu Gly Lys Leu Arg His Glu Asn Val Val Glu Val Val 385 390 395 400 Ala Cys Tyr Phe Ser Lys Asp Glu Lys Leu Val Val Tyr Asp His Val                 405 410 415 Pro Gly Arg Ser Leu Phe His Leu Leu His Glu Asn Arg Gly Glu Gly             420 425 430 Arg Thr Pro Leu Pro Trp Pro Ala Arg Leu Ala Ile Ala Lys Gly Val         435 440 445 Ala Arg Gly Leu Ala Tyr Leu His Gln Thr Leu Pro Leu Phe His Arg     450 455 460 Pro Pro His Gly Asp Leu Lys Ser Ser Asn Val Leu Val Val Phe Pro 465 470 475 480 Gly Pro Gly Gly Arg Gly Gly Gly Gly Asp Ala Val Val Ala                 485 490 495 Lys Leu Thr Asp His Gly Phe His Pro Leu Leu Pro His His His Ala His             500 505 510 Arg Leu Ala Ala Ala Lys Cys Pro Glu Leu Ala Arg Gly Arg Arg Arg         515 520 525 Leu Ser Ser Arg Ala Asp Val Phe Cys Leu Gly Leu Val Leu Leu Glu     530 535 540 Val Val Thr Gly Lys Val Val Val Asp Glu Asp Gly Asp Leu Ala Glu 545 550 555 560 Trp Ala Arg Leu Ala Leu Ser His Glu Trp Ser Thr Asp Ile Leu Asp                 565 570 575 Val Glu Ile Val Ala Asp Arg Gly Arg His Gly Asp Met Leu Arg Leu             580 585 590 Thr Glu Val Ala Leu Leu Cys Ala Ala Val Asp Pro Glu Arg Arg Pro         595 600 605 Lys Ala His Asp Val Val Arg Met Ile Asp Asp Ile Ala Ala Gly Ser     610 615 620 Ala Ala Ala Aly Gly Asp Gly Aly Gla Ala Thr Ala Gly 625 630 635 640 Leu Arg         <210> 2 <211> 16174 <212> DNA <213> Inserting Sequence <400> 2 gtttacccgc caatatatcc tgtcaaacac ggatccgagg taccaggtac caggtgagtt 60 ccattcttac taccacggtg ctattttttt tgctatgtgg ctaattacat gactaacttg 120 gggtgctaaa tcttacaggt tatatgcagg ttatatgcag gtcccgggta ggtcagtccc 180 ttatgttacg tcctgtagaa accccaaccc gtgaaatcaa aaaactcgac ggcctgtggg 240 cattcagtct ggatcgcgaa aactgtggaa ttgatcagcg ttggtgggaa agcgcgttac 300 aagaaagccg ggcaattgct gtgccaggca gttttaacga tcagttcgcc gatgcagata 360 ttcgtaatta tgcgggcaac gtctggtatc agcgcgaagt ctttataccg aaaggttggg 420 caggccagcg tatcgtgctg cgtttcgatg cggtcactca ttacggcaaa gtgtgggtca 480 ataatcagga agtgatggag catcagggcg gctatacgcc atttgaagcc gatgtcacgc 540 cgtatgttat tgccgggaaa agtgtacgta tcaccgtttg tgtgaacaac gaactgaact 600 ggcagactat cccgccggga atggtgatta ccgacgaaaa cggcaagaaa aagcagtctt 660 acttccatga tttctttaac tatgccggaa tccatcgcag cgtaatgctc tacaccacgc 720 cgaacacctg ggtggacgat atcaccgtgg tgacgcatgt cgcgcaagac tgtaaccacg 780 cgtctgttga ctggcaggtg gtggccaatg gtgatgtcag cgttgaactg cgtgatgcgg 840 atcaacaggt ggttgcaact ggacaaggca ctagcgggac tttgcaagtg gtgaatccgc 900 acctctggca accgggtgaa ggttatctct atgaactgtg cgtcacagcc aaaagccaga 960 cagagtgtga tatctacccg cttcgcgtcg gcatccggtc agtggcagtg aagggccaac 1020 agttcctgat taaccacaaa ccgttctact ttactggctt tggtcgtcat gaagatgcgg 1080 acttacgtgg caaaggattc gataacgtgc tgatggtgca cgaccacgca ttaatggact 1140 ggattggggc caactcctac cgtacctcgc attaccctta cgctgaagag atgctcgact 1200 gggcagatga acatggcatc gtggtgattg atgaaactgc tgctgtcggc tttaacctct 1260 ctttaggcat tggtttcgaa gcgggcaaca agccgaaaga actgtacagc gaagaggcag 1320 tcaacgggga aactcagcaa gcgcacttac aggcgattaa agagctgata gcgcgtgaca 1380 aaaaccaccc aagcgtggtg atgtggagta ttgccaacga accggatacc cgtccgcaag 1440 tgcacgggaa tatttcgcca ctggcggaag caacgcgtaa actcgacccg acgcgtccga 1500 tcacctgcgt caatgtaatg ttctgcgacg ctcacaccga taccatcagc gatctctttg 1560 atgtgctgtg cctgaaccgt tattacggat ggtatgtcca aagcggcgat ttggaaacgg 1620 caggaaggt actggaaaaa gaacttctgg cctggcagga gaaactgcat cagccgatta 1680 tcatcaccga atacggcgtg gatacgttag ccgggctgca ctcaatgtac accgacatgt 1740 ggagtgaaga gtatcagtgt gcatggctgg atatgtatca ccgcgtcttt gatcgcgtca 1800 gcgccgtcgt cggtgaacag gtatggaatt tcgccgattt tgcgacctcg caaggcatat 1860 tgcgcgttgg cggtaacaag aaagggatct tcactcgcga ccgcaaaccg aagtcggcgg 1920 cttttctgct gcaaaaacgc tggactggca tgaacttcgg tgaaaaaccg cagcagggag 1980 gcaaacaatg aatcaacaac tctcctggcg caccatcgtc ggctacagcc tcgggaattg 2040 ctaccgagct cgaatttccc cgatcgttca aacatttggc aataaagttt cttaagattg 2100 aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat 2160 gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc 2220 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa 2280 ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat taattcatcg ataggctagt 2340 catggtgact gtacgttgta agtgcagcaa actgccgacg cgatgcaaac tgtacacgtt 2400 aacatgccac tcacctggaa cgcacaatgg ccactaggtg cggccgtagt gtggatttca 2460 aagagagaga gagagagaga gagctaatca cgtaaacgta aacacagcag atagcagaga 2520 tgttgattag gcaaaacagt ataaaagcca atccaataaa ctacatttag cgaagtgcta 2580 tactaatgca ctaataacga actgttcttt tcttaagatc ggagccagta atgggttgtc 2640 agcaggagaa gcacgtaaac cttgaaacat actaagttcc acagtcgaga gtaaaccgta 2700 atcaacacaa gaaacaaaca taaaattgaa caaacgcgca tattataagt gacgaagcgg 2760 tctcacataa aacagggcac acaggttaca acaacgaggg ttgtaagccc attaagcccc 2820 aaacatcaga tcaccacaag caaatgtctc gaagacacac gcacacggca acaggataac 2880 tccacactgg cagatcatgg gatagcagca gttatcaatc aggccttgac acacagaaca 2940 tcaagccccc agacgacgac gactcctcta gatcccggtc ggcatctact ctattccttt 3000 gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca 3060 tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg 3120 gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca 3180 accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc 3240 gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc 3300 aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc 3360 tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg 3420 aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca 3480 tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac 3540 acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct 3600 tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc 3660 atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac 3720 gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg 3780 tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct 3840 ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg 3900 aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg 3960 aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttcatatct 4020 cattgccccc cgggatccgt cgagtcagcc tgaaaggaca aaatacatgt tagcgcctta 4080 gtggtacatt attatttcag tacagcaaga taacacaatt caaaagactg acccataata 4140 aataactagt cctcaattta aaatttgagt tcctaaatag acatctatga atatgctgta 4200 catcggcact acagaaaata cgattcccaa taattgaaca attgtacttt atttagttgt 4260 tactacaaca atggaagata caagatcgtt tcaaaactac catacatgca tggagtattt 4320 gttccacaga tctggaaaaa acagatctga cgggcagtgt caccaaacac tagacatatg 4380 tatttgtatt aggtggatga cgtgtacaaa catgactacc agatctagaa ttagaacgcg 4440 gcgtgctttg gaaatgttaa gtagatccaa atacatcggt aacaaatgaa catattcata 4500 tgacatagct gtaaaacatc atgatctatc atatcaacta gaggggatct cggcatgtat 4560 cattctattg catctagaac catagcattt ccatgtgaca aacaattttg aaacatacat 4620 tgctcagatc taccataaga acaccatgtt catgacagca tcgaccatga tttccacatt 4680 taacagcatc ccgaggtcag atccagtgtt tagaactatc atttcagcaa aattcacaaa 4740 ataaatcctc caattcgcct acatatatcc attccacgca tcctaggacc agatccacca 4800 taccagtata cacgaacact aaacatttca tcagatccgc taactacgac agagaaaacg 4860 cagatccaga caaccagatc caccgacaaa taaacacagc cccccacatc caacaaccgc 4920 gaatccaccc agatctgacc cagaagcagg cacgaagaac acggggggtg agggagatgg 4980 gacgcggatc caagcttggc ggcggattgg gttgatgctg cgacggcggc ggagaaggga 5040 gagaggggag aggaggaaaa gcgcggaggc gcggagagag gcgagtacga agacgccttt 5100 ctctgcgttg tctcggctta gggtttgcga tccccgcact ccgcccgttt tatagggcca 5160 gacagcctgg acctctcagg agcggaagcg aagggatggg ggagtttttc gttttacccc 5220 ctttgacgtc ttcaaaattc actcgcatct cacatcgtgc ctgcaaaggt cgggtggttg 5280 gatcatatcg agaatattta ataagttagg gagctttctg ataatatcca ccgcaaggag 5340 gccatttcgt ttgcactcgg ttgaagtcca tacccgcctc tcgttagttt cactgacagg 5400 tgggtccaga ggcttccttc gtgtgcggtt agcgagtgca cgcgtcgctc caagaaaaag 5460 cagtaggtga cctggccacc tcgttgtagg tgcgaccgat gcagacgtcc cgcttgccgg 5520 tgggcccacg ttacagctgg gtcccacata tcggtgggtg cgatgagttg ttcggtgcgt 5580 aggagtagat tggatcccaa cgggtggttg cgtcgtcggc ggccctgccg attttatttt 5640 cgatttttga aatgcgagag cgggagaacg gcaccgttgg cttggctgtg gatgccgtta 5700 gccgcaccgg actattcggc ccagttccat tttggcccaa cttgaaacag cgcggagcac 5760 aagacggggg agcccattta ggcccgtaat ctcagccctg tagaaagtcc gtgtcgttgc 5820 gatggaccag aaagcccata aaccttggag gcgttctctg ctgggaataa aataagttca 5880 cactctgtcc ctcaacttta cgtcgagttt gtttgacatc gctaatgccc agtaccagaa 5940 atcttggata caaataaggt ttgatataaa gtggcacgtg gtgaatggca tcgtttgaaa 6000 aatcttcaga tcgagcgggc ttcacaagcc gtaagtgcaa gtgccaaaat ttgtgaagaa 6060 aattacttcc ttcatttcac aatgtaagtt attttagcat tttccacatt catattgata 6120 tcaataaatt tagatagata tatatgtctg aattcgtacc gttaacgagc tctagaagct 6180 tcagtactac gcgtctcgag gcctattatt gaagcattta tcagggttat tgtctcatga 6240 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6300 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa 6360 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct 6420 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac 6480 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg 6540 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg 6600 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag 6660 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 6720 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 6780 gtgaattcac tagtgattgc tctagacact gatagtttcg gatctagata tcacatcaat 6840 ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgttc ctcgtgggtg 6900 ggggtccatc tttgggacca ctgtcggtag aggcatcttg aacgatagcc tttcctttat 6960 cgcaatgatg gcatttgtag aagccatctt ccttttctac tgtcctttcg atgaagtgac 7020 agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct 7080 caatagccct ctggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt 7140 cgtgctccac catgttgggg atctagatat cacatcaatc cacttgcttt gaagacgtgg 7200 ttggaacgtc ttctttttcc acgatgttcc tcgtgggtgg gggtccatct ttgggaccac 7260 tgtcggtaga ggcatcttga acgatagcct ttcctttatc gcaatgatgg catttgtaga 7320 agccatcttc cttttctact gtcctttcga tgaagtgaca gatagctggg caatggaatc 7380 cgaggaggtt tcccgatatt accctttgtt gaaaagtctc aatagccctc tggtcttctg 7440 agactgtatc tttgatattc ttggagtaga cgagagtgtc gtgctccacc atgttgggga 7500 tctagatatc acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca 7560 cgatgttcct cgtgggtggg ggtccatctt tgggaccact gtcggtagag gcatcttgaa 7620 cgatagcctt tcctttatcg caatgatggc atttgtagaa gccatcttcc ttttctactg 7680 tcctttcgat gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta 7740 ccctttgttg aaaagtctca atagccctct ggtcttctga gactgtatct ttgatattct 7800 tggagtagac gagagtgtcg tgctccacca tgttggggat ctagatatca catcaatcca 7860 cttgctttga agacgtggtt ggaacgtctt ctttttccac gatgttcctc gtgggtgggg 7920 gtccatcttt gggaccactg tcggtagagg catcttgaac gatagccttt cctttatcgc 7980 aatgatggca tttgtagaag ccatcttcct tttctactgt cctttcgatg aagtgacaga 8040 tagctgggca atggaatccg aggaggtttc ccgatattac cctttgttga aaagtctcaa 8100 tagccctctg gtcttctgag actgtatctt tgatattctt ggagtagacg agagtgtcgt 8160 gctccaccat gttggggatc ctctagagtc gagaattcag tacattaaaa acgtccgcaa 8220 tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc tgccaccagc 8280 cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata caggcagccc 8340 atcagtccgg gacggcgtca gcgggagagc cgttgtaagg cggcagactt tgctcatgtt 8400 accgatgcta ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg gatgatctcg 8460 cggagggtag catgttgatt gtaacgatga cagagcgttg ctgcctgtga tcaaatatca 8520 tctccctcgc agagatccga attatcagcc ttcttattca tttctcgctt aaccgtgaca 8580 ggctgtcgat cttgagaact atgccgacat aataggaaat cgctggataa agccgctgag 8640 gaagctgagt ggcgctattt ctttagaagt gaacgttgac gatcgtcgac ggatcttttc 8700 cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt 8760 cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg 8820 gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac 8880 tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccggc 8940 taccgccggc gtaacagatg agggcaagcg gatggctgat gaaaccaagc caaccaggaa 9000 gggcagccca cctatcaagg tgtactgcct tccagacgaa cgaagagcga ttgaggaaaa 9060 ggcggcggcg gccggcatga gcctgtcggc ctacctgctg gccgtcggcc agggctacaa 9120 aatcacgggc gtcgtggact atgagcacgt ccgcgagctg gcccgcatca atggcgacct 9180 gggccgcctg ggcggcctgc tgaaactctg gctcaccgac gacccgcgca cggcgcggtt 9240 cggtgatgcc acgatcctcg ccctgctggc gaagatcgaa gagaagcagg acgagcttgg 9300 caaggtcatg atgggcgtgg tccgcccgag ggcagagcca tgactttttt agccgctaaa 9360 acggccgggg ggtgcgcgtg attgccaagc acgtccccat gcgctccatc aagaagagcg 9420 acttcgcgga gctggtattc gtgcagggca agattcggaa taccaagtac gagaaggacg 9480 gccagacggt ctacgggacc gacttcattg ccgataaggt ggattatctg gacaccaagg 9540 caccaggcgg gtcaaatcag gaataagggc acattgcccc ggcgtgagtc ggggcaatcc 9600 cgcaaggagg gtgaatgaat cggacgtttg accggaaggc atacaggcaa gaactgatcg 9660 acgcggggtt ttccgccgag gatgccgaaa ccatcgcaag ccgcaccgtc atgcgtgcgc 9720 cccgcgaaac cttccagtcc gtcggctcga tggtccagca agctacggcc aagatcgagc 9780 gcgacagcgt gcaactggct ccccctgccc tgcccgcgcc atcggccgcc gtggagcgtt 9840 cgcgtcgtct cgaacaggag gcggcaggtt tggcgaagtc gatgaccatc gacacgcgag 9900 gaactatgac gaccaagaag cgaaaaaccg ccggcgagga cctggcaaaa caggtcagcg 9960 aggccaagca ggccgcgttg ctgaaacaca cgaagcagca gatcaaggaa atgcagcttt 10020 ccttgttcga tattgcgccg tggccggaca cgatgcgagc gatgccaaac gacacggccc 10080 gctctgccct gttcaccacg cgcaacaaga aaatcccgcg cgaggcgctg caaaacaagg 10140 tcattttcca cgtcaacaag gacgtgaaga tcacctacac cggcgtcgag ctgcgggccg 10200 acgatgacga actggtgtgg cagcaggtgt tggagtacgc gaagcgcacc cctatcggcg 10260 agccgatcac cttcacgttc tacgagcttt gccaggacct gggctggtcg atcaatggcc 10320 ggtattacac gaaggccgag gaatgcctgt cgcgcctaca ggcgacggcg atgggcttca 10380 cgtccgaccg cgttgggcac ctggaatcgg tgtcgctgct gcaccgcttc cgcgtcctgg 10440 accgtggcaa gaaaacgtcc cgttgccagg tcctgatcga cgaggaaatc gtcgtgctgt 10500 ttgctggcga ccactacacg aaattcatat gggagaagta ccgcaagctg tcgccgacgg 10560 cccgacggat gttcgactat ttcagctcgc accgggagcc gtacccgctc aagctggaaa 10620 ccttccgcct catgtgcgga tcggattcca cccgcgtgaa gaagtggcgc gagcaggtcg 10680 gcgaagcctg cgaagagttg cgaggcagcg gcctggtgga acacgcctgg gtcaatgatg 10740 acctggtgca ttgcaaacgc tagggccttg tggggtcagt tccggctggg ggttcagcag 10800 ccagcgcttt actggcattt caggaacaag cgggcactgc tcgacgcact tgcttcgctc 10860 agtatcgctc gggacgcacg gcgcgctcta cgaactgccg ataaacagag gattaaaatt 10920 gacaattgtg attaaggctc agattcgacg gcttggagcg gccgacgtgc aggatttccg 10980 cgagatccga ttgtcggccc tgaagaaagc tccagagatg ttcgggtccg tttacgagca 11040 cgaggagaaa aagcccatgg aggcgttcgc tgaacggttg cgagatgccg tggcattcgg 11100 cgcctacatc gacggcgaga tcattgggct gtcggtcttc aaacaggagg acggccccaa 11160 ggacgctcac aaggcgcatc tgtccggcgt tttcgtggag cccgaacagc gaggccgagg 11220 ggtcgccggt atgctgctgc gggcgttgcc ggcgggttta ttgctcgtga tgatcgtccg 11280 acagattcca acgggaatct ggtggatgcg catcttcatc ctcggcgcac ttaatatttc 11340 gctattctgg agcttgttgt ttatttcggt ctaccgcctg ccgggcgggg tcgcggcgac 11400 ggtaggcgct gtgcagccgc tgatggtcgt gttcatctct gccgctctgc taggtagccc 11460 gatacgattg atggcggtcc tgggggctat ttgcggaact gcgggcgtgg cgctgttggt 11520 gttgacacca aacgcagcgc tagatcctgt cggcgtcgca gcgggcctgg cgggggcggt 11580 ttccatggcg ttcggaaccg tgctgacccg caagtggcaa cctcccgtgc ctctgctcac 11640 ctttaccgcc tggcaactgg cggccggagg acttctgctc gttccagtag ctttagtgtt 11700 tgatccgcca atcccgatgc ctacaggaac caatgttctc ggcctggcgt ggctcggcct 11760 gatcggagcg ggtttaacct acttcctttg gttccggggg atctcgcgac tcgaacctac 11820 agttgtttcc ttactgggct ttctcagccc ggggaccgcc gtgttgctag gatggttgtt 11880 cttggatcag acgctgagtg cgcttcaaat catcggcgtc ctgctcgtga tcgggagtat 11940 ctggctgggc caacgttcca accgcactcc tagggcgcgt atagcttgcc ggaagtcgcc 12000 ttgacccgca tggcataggc ctatcgtttc cacgatcagc gatcggctcg ttgccctgcg 12060 ccgctccaaa gcccgcgacg cagcgccggc aggcagagca agtagagggc agcgcctgca 12120 atccatgccc acccgttcca cgttgttata gaagccgcat agatcgccgt gaagaggagg 12180 ggtccgacga tcgaggtcag gctggtgagc gccgccagtg agccttgcag ctgcccctga 12240 cgttcctcat ccacctgcct ggacaacatt gcttgcagcg ccggcattcc gatgccaccc 12300 gaagcaagca ggaccatgat cgggaacgcc atccatcccc gtgtcgcgaa ggcaagcagg 12360 atgtagcctg tgccgtcggc aatcattccg agcatgagtg cccgcctttc gccgagccgg 12420 gcggctacag ggccggtgat cattgcctgg gcgagtgaat gcagaatgcc aaatgcggca 12480 agcgaaatgc cgatcgtggt cgcgtcccag tgaaagcgat cctcgccgaa aatgacccaa 12540 agcgcggccg gcacctgtcc gacaagttgc atgatgaaga agaccgccat cagggcggcg 12600 acgacggtca tgccccgggc ccaccggaac gaagcgagcg ggttgagagc ctcccggcgt 12660 aacggccggc gttcgccttt gtgcgactcc ggcaaaagga aacagcccgt caggaaattg 12720 aggccgttca aggctgccgc ggcgaagaac ggagcgtggg gggagaaacc gcccatcagc 12780 ccaccgagca caggtcccgc gaccatcccg aacccgaaac aggcgctcat gaagccgaag 12840 tgccgcgcgc gctcatcgcc atcagtgata tcggcaatat aagcgccggc taccgcccca 12900 gtcgccccgg tgatgccggc cacgatccgc ccgatataga gaacccaaag gaaaggcgct 12960 gtcgccatga tggcgtagtc gacagtggcg ccggccagcg agacgagcaa gattggccgc 13020 cgcccgaaac gatccgacag cgcgcccagc acaggtgcgc aggcaaattg caccaacgca 13080 tacagcgcca gcagaatgcc atagtgggcg gtgacgtcgt tcgagtgaac cagatcgcgc 13140 aggaggcccg gcagcaccgg cataatcagg ccgatgccga cagcgtcgag cgcgacagtg 13200 ctcagaatta cgatcagggg tatgttgggt ttcatgtctg gcctccggac cagcctccgc 13260 tggtccgatt gaacgcgcgg attctttatc actgataagt tggtggacat attatgttta 13320 tcagtgataa agtgtcaagc atgacaaagt tgcagccgaa tacagtgatc cgtgccgccc 13380 tggacctgtt gaacgaggtc ggcgtagacg gtctgacgac acgcaaactg gcggaacggt 13440 tgggggttca gcagccggcg ctttactggc acttcaggaa caagcgggcg ctgctcgacg 13500 cactggccga agccatgctg gcggagaatc atacgcattc ggtgccgaga gccgacgacg 13560 actggcgctc atttctgatc gggaatgccc gcagcttcag gcaggcgctg ctcgcctacc 13620 gcgatggcgc gcgcatccat gccggcacgc gaccgggcgc accgcagatg gaaacggccg 13680 acgcgcagct tcgcttcctc tgcgaggcgg gtttttcggc cggggacgcc gtcaatgcgc 13740 tgatgacaat cagctacttc actgttgggg ccgtgcttga ggagcaggcc ggcgacagcg 13800 atgccggcga gcgcggcggc accgttgaac aggctccgct ctcgccgctg ttgcgggccg 13860 cgatagacgc cttcgacgaa gccggtccgg acgcagcgtt cgagcaggga ctcgcggtga 13920 ttgtcgatgg attggcgaaa aggaggctcg ttgtcaggaa cgttgaagga ccgagaaagg 13980 gtgacgattg atcaggaccg ctgccggagc gcaacccact cactacagca gagccatgta 14040 gacaacatcc cctccccctt tccaccgcgt cagacgcccg tagcagcccg ctacgggctt 14100 tttcatgccc tgccctagcg tccaagcctc acggccgcgc tcggcctctc tggcggcctt 14160 ctggcgctcc tgctgcggcg tccgctcgtg ggccgtggcg cgggtccgcg cgccggcctc 14220 gtgcgcctgg cgctcgcggg cgaggtccag ggcggccgtc ttcacgttct gccttgcgca 14280 gatgagatag atccagtctg caagtgtcag aaacgagtct gcatttgatg ccctgaacgt 14340 cttcatacat ctgataagcc cctcccggaa gtctgctttc tctccgtcag aaagcatacc 14400 ctgaccggtt tcaatcacgg accgttctgt caggtaatgc tctgtccagt catacagagc 14460 cgggtttgtc acccgtataa gctgaagcag gcacaaatca gggaaataaa caaaatcccg 14520 catccccgga taaagaaaaa tcagggaatt aatggcctga tggatttccc gtggcgtcga 14580 aagtgcggca ccataggtat cagtcaccgc gatgagatcc cttaccattc cagagtctgg 14640 cggttgatta ttaatttgct gatatagagc ctcagcccgc tggcgaaatt cattacgtaa 14700 atcaaaggct tcaggtcggg gtaatttaaa actaagctga atgattttct ggagataacg 14760 gctgccatct tcgatattca gcgcatgttc aacggcatga gtgataatct gcctgtcata 14820 acagagaata tgggtaaagc ggggcagatc ggctactgca cgcacaagcc tgaacacttc 14880 cgccacctgg gatggctcca gtcggtccag atcatccatg acaacaatga acttcagatc 14940 cagactcacc agttgtcctg caatttcagc ccgaagctag cttggctgcc atttttgggg 15000 tgaggccgtt cgcggccgag gggcgcagcc cctgggggga tgggaggccc gcgttagcgg 15060 gccgggaggg ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca cgcgcacagg 15120 gcgcagccct ggttaaaaac aaggtttata aatattggtt taaaagcagg ttaaaagaca 15180 ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt tctgcctgtg 15240 gacagcccct caaatgtcaa taggtgcgcc cctcatctgt cagcactctg cccctcaagt 15300 gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca ataccgcagg 15360 gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa tcaggcgttt 15420 tcgccgattt gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc ctgcccctca 15480 tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc ccctcatctg 15540 tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc ggcggccgcg gtgtctcgca 15600 cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc cagcccagcg 15660 gcgagggcaa ccagcccggt gagcgtcgga aagggtcgac gatcttgctg cgttcggata 15720 ttttcgtgga gttcccgcca cagacccgga ttgaaggcga gatccagcaa ctcgcgccag 15780 atcatcctgt gacggaactt tggcgcgtga tgactggcca ggacgtcggc cgaaagagcg 15840 acaagcagat cacgcttttc gacagcgtcg gatttgcgat cgaggatttt tcggcgctgc 15900 gctacgtccg cgaccgcgtt gagggatcaa gccacagcag cccactcgac cttctagccg 15960 acccagacga gccaagggat ctttttggaa tgctgctccg tcgtcaggct ttccgacgtt 16020 tgggtggttg aacagaagtc attatcgcac ggaatgccaa gcactcccga ggggaaccct 16080 gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc cttttaaata 16140 tccgattatt ctaataaacg ctcttttctc ttag 16174 <210> 3 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> primer Enhancer-F <400> 3 ccggtaccat ggcggcggtg gtggaggg 28 <210> 4 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer Enhancer-R <400> 4 ccggatcctt agcttagaga tgagacgaa 29 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer pGA2715L1.5S-F <400> 5 agtacattaa aaacgtccgc 20 <210> 6 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> primer rt-Os10g35040-F <400> 6 gcgctgagcc acgagt 16 <210> 7 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> primer rt-Os10g35040-R <400> 7 gtcgcctccc cgtctc 16 <210> 8 <211> 1929 <212> DNA <213> Oryza sativa LOC_Os10g35040 <400> 8 atggcggcgg tggtggaggg tgtggggtgg ccagagccgg agctggaggc gttgcgcgac 60 gagcggggcg ggctggtggc gctgcgggac gcgctgcggt cggggaggga cctgcactcg 120 aactggacgg ggccgccgtg ccacggcggg cggagccggt ggtacggcgt ggcgtgcgac 180 ggcgacgggc gggtggtcgg cgtgcagctc gacggcgcgc agctcacggg cgcgctcccc 240 gcgggggcgc tcgccggcgt ggcgcggctg gagacgctgt ccctgcgcga caacgccatc 300 ccggcgcgc tcccgcggct cgacgcgctg gcgcggctcc gcgtcgtgga cctgtcgagc 360 aaccggttct cggggcccat cccgcgcggg tacgcggcgg cgctgggtga gctcacgcgg 420 ctcgagctcc aggacaacct gatcaatggc acgctgccgg cgttcgagca ggacgggctc 480 gccgtgttca acgtgtcgta caacttcctc cagggcgagg tgcccgacac ccgcgcgctc 540 cgccggttcc ccgccaccgc gttcgcccac aacctcaggc tctgcggcga ggttgtgagg 600 acggagtgcc gccgcgaggg ctcgccattc gacgccgcgc cggccggcgg cggcggcagc 660 ggcagcgatg gtggtgatcg ggtcttcggg gcgcgcgacg ccgccgcgcc gccggcgcgg 720 tggcgcaagc ccatccggtt caggatagcg aggtggagcg tcgtggtgat cgcgctcatc 780 gccgcgctgg tgccgttcgc cgcggtgctc atcttcctcc accacagcaa gaagagccgg 840 gtggacaagg cggcggagca agccggcaag aaggtgagca gtggcagcgg caacggaagc 900 cggagcacga cggagtccgg caagggagcg gcggatcagc tacagttctt ccggccggag 960 aaggcgacct tcagcctcga cgagctgttc cgttcgacgg cggagatgct cggcaagggc 1020 cggctaggga tcacctaccg cgtcgccctc cacgccggcg gcggcggcgg cggcggcggc 1080 ggcccggtcg tcgtcgtcgt gaagcggctg cgcaacatgg ggcacgtgcc acgcaaggac 1140 ttcgcccaca ccatgcagct cctcggcaag ctccgccatg agaacgtcgt cgaggtcgtc 1200 gcctgctact tctccaagga cgagaagctc gtcgtctacg accacgtccc cggccgcagc 1260 ctcttccacc tcctccacga gaacagagga gaggggagga cgccgctgcc atggccggcg 1320 aggttggcga tagcgaaggg cgtggcgcgg gggctggcct acctgcacca gacgctgccg 1380 ctcttccacc ggccgccgca cggcgacctc aagtcgtcga acgtgctcgt cgtcttcccg 1440 gggcccggcg gccgcggagg cggcggcggc gacgcggtgc cggtggccaa gctgactgac 1500 cacgggttcc acccgctgct gccgcaccac gcgcaccgcc tcgcggcggc caagtgcccg 1560 ggctcgcgc gcgggaggcg gcggctctcc tcccgcgccg acgtgttctg cctcggcctc 1620 gtgctgctcg aggtggtcac cgggaaggtc cccgtcgacg aggacggcga cctggcggag 1680 tgggcgcggc tggcgctgag ccacgagtgg tcgacggaca tcctcgacgt ggagatcgtc 1740 gccgaccggg gccgccacgg cgacatgctg aggctcaccg aggtcgccct gctctgcgcc 1800 gccgtcgacc ccgagcgccg gcccaaggcg cacgacgtcg tcaggatgat cgacgacatc 1860 gccgccggct ccgccgccgc cgccggagac ggggaggcga cggcggggcg agagctcgcg 1920 ctgcggtga 1929 <210> 9 <211> 1400 <212> DNA <213> Enhancing Sequence <400> 9 gaattgatcc ccaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat 60 acagtctcag aagaccagag ggctattgag acttttcaac aaagggtaat atcgggaaac 120 ctcctcggat tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa 180 gatggcttct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct 240 accgacagtg gtcccaaaga tggaccccca cccacgagga acatcgtgga aaaagaagac 300 gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctagatccc caacatggtg 360 gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga agaccagagg 420 gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt ccattgccca 480 gctatctgtc acttcatcga aaggacagta gaaaaggaag atggcttcta caaatgccat 540 cattgcgata aaggaaaggc tatcgttcaa gatgcctcta ccgacagtgg tcccaaagat 600 ggacccccac ccacgaggaa catcgtggaa aaagaagacg ttccaaccac gtcttcaaag 660 caagtggatt gatgtgatat ctagatcccc aacatggtgg agcacgacac tctcgtctac 720 tccaagaata tcaaagatac agtctcagaa gaccagaggg ctattgagac ttttcaacaa 780 agggtaatat cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcgaa 840 aggacagtag aaaaggaaga tggcttctac aaatgccatc attgcgataa aggaaaggct 900 atcgttcaag atgcctctac cgacagtggt cccaaagatg gacccccacc cacgaggaac 960 atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc 1020 tagatcccca acatggtgga gcacgacact ctcgtctact ccaagaatat caaagataca 1080 gtctcagaag accagagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc 1140 ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga aaaggaagat 1200 ggcttctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga tgcctctacc 1260 gacagtggtc ccaaagatgg acccccaccc acgaggaaca tcgtggaaaa agaagacgtt 1320 ccaaccacgt cttcaaagca agtggattga tgtgatatct agatccgaaa ctatcagtgt 1380 ctagctagag tcgagaattc 1400

Claims (15)

As the transformed rice (Oryza sp.) In which LOC_Os10g35040, an inherent inactive gene consisting of the nucleotide sequence shown in SEQ ID NO: 8, is activated,
Wherein the rice comprises an expression inducing sequence activating LOC_Os10g35040 or a recombinant vector for promoting resistance to blight of blight of rice into which a promoter has been inserted.
delete delete The transformed rice according to claim 1, wherein the rice (Oryza sp.) Is Dongjinbyeong or Kwangjungpyeong.
delete The transformed rice of claim 1, wherein the expression inducing sequence comprises the sequence as set forth in SEQ ID NO: 9.
7. The transformed rice according to claim 6, wherein the expression inducing sequence is a nucleotide sequence as set forth in SEQ ID NO: 2.
delete (a) introducing a recombinant vector for transformation into an Agrobacterium containing an expression inducing sequence for expressing LOC_Os10g35040, which is an inherent inactive gene consisting of the nucleotide sequence shown in SEQ ID NO: 8;
(b) infecting the rice leaves with the Agrobacterium of step (a); And
(c) callusing the rice cells infected with the step (b) to obtain transformed rice cells, the method comprising the steps of:
The disease resistance is resistance to blight of blight,
Wherein said transformant is rice (Oryza sp.).
The method according to claim 9, further comprising: (d) inducing root and adult differentiation of the callus produced in step (c).
delete delete 10. The method according to claim 9, wherein the rice is Dongjinbyeon or Kwangyoungbyeon.
10. The method according to claim 9, wherein the expression inducing sequence of step (a) comprises the nucleotide sequence shown in SEQ ID NO: 9.
8. A method for promoting rice disease resistance comprising activating a LOC_Os10g35040 gene comprising the nucleotide sequence of SEQ ID NO: 8,
The activation is achieved by inserting an expression inducing sequence or a promoter into the LOC_OslOg35040 expression control region,
Wherein said disease resistance is resistance to blight of blight.

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