一种受抗生素诱导的启动子的构建及活性分析

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。

Construction and Activity Analysis of an Antibiotic-inducible Promoter

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.