生物技术通报2017，Vol.33Issue(2)：149-154,6.DOI:10.13560/j.cnki.biotech.bull.1985.2017.02.022
基于四环素调控系统的叶绿体启动子在大肠杆菌中活性检测
Activity Detection of Chloroplast Promoter Based on Tetracycline Regulatory System in Escherichia coli
摘要
Abstract
In order to study tetracycline regulatory system regulating the activity of promoter in chloroplast genetic engineering,having the tetracycline (Tet) gene and the core regulatory area of TetR (Tet repressor) specific recognition sequences as the basis,the promoter prrnO 1O2 with tetracycline core regulatory area was synthetized,then ligated to expression vector Bio3-GFP,and the activity of the promoter was validated in Escherichia coli.The expression of the GFP gene driven by the promoter led colonies to be bright green.The expression vector Bio3-TetR-prmO1O2-GFP for GFP gene induced by Tet was constructed,and screening experiment confirmed that the highest concentration of tetracycline for the proper growth of E.coli was 5 μg/mL.Finally,GFP expression vector based on tetracycline regulatory system was constructed,the function of prrnO1O2 promoter was suppressed while without adding tetracycline.GFP gene expressed to be green fluorescent protein after adding tetracycline.These results indicated that tetracycline regulatory system controlled the activity of chloroplasts promoter prrnO1O2 in E.coli,thus avoiding the regulation interference of nuclear genome to plastid genome,and which provides effective ways and methods to plastid genetic engineering breeding further.关键词
四环素调控系统/叶绿体/启动子/prrnO1O2/大肠杆菌Key words
tetracycline regulatory system/chloroplast/promoter/prrnO1O2/Escherichia coli引用本文复制引用
于晓俊,曹绍玉,董玉梅,毕保良,张应华,许俊强..基于四环素调控系统的叶绿体启动子在大肠杆菌中活性检测[J].生物技术通报,2017,33(2):149-154,6.基金项目
国家自然科学基金项目(31560560,30972012),云南省应用基础研究计划项目(2015FD019) （31560560,30972012）
