CAJ | 학술논문

TGA转录因子通过与NPR1基因协同作用参与植物对病害的防御作用。从水稻突变体HX-3基因组中分离到一个TGA转录因子rTGA4的5′非翻译区1995 bp的序列（ pTGA），该序列与日本晴基因组序列仅有94％的相似性。经PLACE和PlantCARE序列分析表明：该片段含有典型的TATA-box、CAAT-box等基本转录元件，以及脱落酸、乙烯、茉莉酸甲酯、赤霉素以及病原菌响应元件等。将得到的pTGA利用T/A克隆法连接到植物表达载体pCXGUS-T/A上，通过花序浸染法转化拟南芥，并对转基因拟南芥进行分子检测及GUS组织化学染色。结果表明，在苗期时GUS主要在幼苗根尖表达，在其他部位均没有表达；而在成熟期GUS在多处均有表达，特异性并不明显，表明该启动子是受生长发育阶段调控的组织特异性启动子。通过对rTGA4启动子的特征研究，为进一步克隆HX-3中的抗性基因以及利用奠定基础。
TGA전록인자통과여NPR1기인협동작용삼여식물대병해적방어작용。종수도돌변체HX-3기인조중분리도일개TGA전록인자rTGA4적5′비번역구1995 bp적서렬（ pTGA），해서렬여일본청기인조서렬부유94％적상사성。경PLACE화PlantCARE서렬분석표명：해편단함유전형적TATA-box、CAAT-box등기본전록원건，이급탈락산、을희、말리산갑지、적매소이급병원균향응원건등。장득도적pTGA이용T/A극륭법련접도식물표체재체pCXGUS-T/A상，통과화서침염법전화의남개，병대전기인의남개진행분자검측급GUS조직화학염색。결과표명，재묘기시GUS주요재유묘근첨표체，재기타부위균몰유표체；이재성숙기GUS재다처균유표체，특이성병불명현，표명해계동자시수생장발육계단조공적조직특이성계동자。통과대rTGA4계동자적특정연구，위진일보극륭HX-3중적항성기인이급이용전정기출。
The family of TGA transcription factors cooperates with NPR1 to play a very important role in dis-ease defence.A 1 995 bp sequence of the 5′UTR of a new TGA transcription factor rTGA4 was isolated from rice mutant HX-3 genomic DNA and named pTGA ,which shows 94%similarity to the genomic sequence of nipponbare . Sequence analysis of the promoter by PLACE and Plant CARE showed that the cloned fragment contained such some basic transcription element TATA and CAAT-Box, and some putative cis-elements, such as abscisic acid respon-seive,ethylene responsive ,MeJA responsive ,gibberellin-responsive and pathogen responsive elements .Then an ex-pression vector was constructed by connecting the pTGA with pCXGUS-T/A and it was transformed to Arabidopsis thaliana by Agrobacterium-mediated method .The transgenic plants were analyzed by PCR amplification and GUS staining .We found that GUS gene was mainly expressed in shoot tip of seedling and did not detect GUS activity in other parts,but the specific expression was lost during mature period .So we consider that the pTGA is a tissue spe-cific promoter which is regulated by growth stage .Characterization of the promoter of the rTGA 4 lays a foundation for cloning the resistant gene in HX-3 and its further application .