一种受抗生素诱导的启动子的构建及活性分析

doi:10.3724/SP.J.1259.2011.00560

植物学报 ›› 2011, Vol. 46 ›› Issue (5): 560-568.DOI: 10.3724/SP.J.1259.2011.00560

一种受抗生素诱导的启动子的构建及活性分析

刘伟伟, 张海磊, 刘财丰, 葛晓春^*

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

收稿日期:2011-01-31 修回日期:2011-05-13 出版日期:2011-09-01 发布日期:2011-09-01
通讯作者: 葛晓春
基金资助:
国家自然科学基金青年基金项目;国家自然科学基金青年基金项目

Construction and Activity Analysis of an Antibiotic-inducible Promoter

Weiwei Liu, Hailei Zhang, Caifeng Liu, Xiaochun Ge^*

Department of Biochemistry and Molecular Biology, Institute of Plant Science, School of Life Sciences, Fudan University, Shanghai 200433, China

Received:2011-01-31 Revised:2011-05-13 Online:2011-09-01 Published:2011-09-01
Contact: Xiaochun Ge

摘要/Abstract

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2 179 bp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; 1.5 μg·mL^–1博莱霉素与22 μg·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1 μg·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。

Abstract: 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.

刘伟伟, 张海磊, 刘财丰, 葛晓春. 一种受抗生素诱导的启动子的构建及活性分析. 植物学报, 2011, 46(5): 560-568.

Weiwei Liu, Hailei Zhang, Caifeng Liu, Xiaochun Ge. Construction and Activity Analysis of an Antibiotic-inducible Promoter. Chinese Bulletin of Botany, 2011, 46(5): 560-568.

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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

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