一种受抗生素诱导的启动子的构建及活性分析  

Construction and Activity Analysis of an Antibiotic-inducible Promoter

作　　者：刘伟伟[1] 张海磊[1] 刘财丰[1] 葛晓春[1] 

机构地区：[1]复旦大学生命科学学院植物研究所生物化学与分子生物学系,上海200433

出　　处：《植物学报》2011年第5期560-568,共9页Chinese Bulletin of Botany

基　　金：国家自然科学基金(No.30770425和No.31070232)

摘　　要：多聚ADP核糖聚合酶(PARP)受基因毒剂的特异性诱导。将拟南芥(Arabidopsis thaliana)AtPARP1基因上游长2179bp的启动子片段插入到质粒pAKK687的β-葡萄糖醛酸糖苷酶(GUS)报告基因上游,转化拟南芥。GUS组织化学染色结果表明,GUS报告基因仅在苗龄3-5天的拟南芥根部及花发育早期的雄蕊中表达;1.5μg.mL-1博莱霉素与22μg.mL-1丝裂霉素联用强烈诱导了GUS报告基因的表达(尤其在拟南芥的幼苗和果荚中)。进一步降低抗生素浓度,发现单独使用1μg.mL-1博莱霉素对GUS报告基因也具较强的诱导活性,且对拟南芥幼苗的生长无影响。上述结果表明,AtPARP1启动子是一个新型的具较大应用潜力的抗生素诱导型启动子。Poly ADP-ribose polymerase（PARP） is induced specifically by genotoxin.We cloned the 2 179 bp promoter sequence of Arabidopsis AtPARP1 gene into a pAKK687 vector to drive the expression of a β-glucuronidase（GUS） reporter gene for Arabidopsis transformation.GUS staining revealed the GUS reporter gene expressed only in the basal root of 3 to 5-day-old seedlings and in the anther at early flowering stage.An amount of 1.5 μg.mL-1 bleomycin combined with 22 μg.mL-1 mitomycin could strongly induce the expression of the GUS reporter gene,especially in young seedlings and young siliques of Arabidopsis.An amount of 1 μg.mL-1 bleomycin was enough to induce the promoter and had no adverse effects on Arabidopsis seedling growth.The AtPARP1 promoter may be a new antibiotic-inducible promoter with application potential.

关 键 词：抗生素 拟南芥 博莱霉素 诱导型启动子 多聚ADP核糖聚合酶 

分 类 号：Q943.2[生物学—植物学]