检测致畸物的gfp基因工程菌的构建  

Construction of a Genotoxic Detectable GEM Strain with gfp

作　　者：顾立锋[1] 蒋建东[1] 何健[1] 黄星[1] 贾开志[1] 李顺鹏[1] 

机构地区：[1]南京农业大学生命科学学院微生物学系/农业部农业环境微生物工程重点开放实验室,南京210095

出　　处：《应用与环境生物学报》2007年第1期83-86,共4页Chinese Journal of Applied and Environmental Biology

基　　金：国家"863"项目(2003AA241150);江苏省科技攻关项目(BE2003345;BE2003343)~~

摘　　要：RecA与UvrA是细胞SOS系统中重要的修复蛋白,在细胞DNA损伤后诱导表达.将报告基因绿色荧光蛋白基因(gfpmut3a)分别置于recA和uvrA启动子调控下,构建成质粒pRecAgfp和pUvrAgfp,并转化E.coliDH5α,构建成检测菌株ERS101和EUS101.试验发现,菌株ERS101和EUS101在致畸物吖啶橙处理后,GFP的表达量增高,菌悬液经507nm的蓝光激发后产生的绿色荧光明显增强,且与吖啶橙的浓度具有一定的相关性,即使在0.5μgmL-1的吖啶橙诱导下也能检测到荧光增强效应.对硝基酚等多种致畸物处理后的检测菌株均有荧光增强效应.RecA or UvrA was one of the most important DNA repair proteins in SOS system. Plasmids pRecAgfp and pUvrAgfp were constructed after DNA damage-inducible promoters of recA and uvrA from Escherichia co/i were fused to the reporter gene gfpmut3a operon. Genotoxic detectable Strains ERS101 and EUS101 were obtained after introducing plasmids pRecAgfp or pUvrAgfp into E. coli DH5α. Strains ERS101 and EUS101 produced More bioluminescent when treated with genotoxic acridine orange even at 0.5μg mL^-1 concentrations, and the bioluminescent intensity was correlated with the concentration of acridine orange, more bioluminescent was also detected when treated with other genotoxic such as 4-nitrophenol. Fig 4, Tab 1, Ref 9 .

关 键 词：GFP 致畸物 基因工程菌构建 

分 类 号：Q789[生物学—分子生物学]