绿色荧光蛋白基因在枯草芽孢杆菌中的表达  

Expression of Green Fluorescent Protein Gene in Bacillus subtilis

作　　者：陈晓月[1] 金宁一[1] 海洋[2] 邹伟[2] 马海利[1] 李昌[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]军事医学科学院十一所全军基因工程重点实验室

出　　处：《中国生物制品学杂志》2008年第2期115-118,共4页Chinese Journal of Biologicals

基　　金："863"计划项目(2006AA10A205);教育部重点项目(306006);吉林省重点项目(20065020)

摘　　要：目的探讨异源基因在枯草芽孢杆菌中表达的可行性。方法以大肠杆菌和枯草芽孢杆菌的穿梭质粒为基本骨架,在大肠杆菌中完成含有绿色荧光蛋白(GFP)基因的重组表达质粒的构建,通过酶切鉴定筛选阳性克隆,采用电转化的方法,将重组质粒转入枯草芽孢杆菌进行表达,通过检测GFP的存在,评价枯草芽孢杆菌对异源基因的表达。结果重组表达质粒pGJP-GFP酶切鉴定结果与预期片段大小相符。绿色荧光蛋白可在枯草芽孢杆菌中表达,且表达蛋白具有较好的生物学活性。结论利用枯草芽孢杆菌的自身启动子,可以实现外源基因在枯草芽孢杆菌中的表达。Objective To explore the feasibihty of expressing foreign gene in Bacillus subtilis. Methods Construct a recombinant plasmid containing green fluorescent protein （GFP） gene based on a shuttle vector pGj103 of E. coli and B. subtilis and identify by restriction analysis. Screen positive clones and transform to B. subtilis by electrotransformation for expression. Determine the transformed B. subtilis for presence of GFP to evaluate the expression of foreign gene. Results The restriction analysis of recombinant expression vector pGJP-GFP proved that the lengths of obtained gene fragments were consistent with those expected. GFP was successfidly expressed in B. subtilis and showed good biological activity. Conclusion It is feasible to express foreign gene in B. subtilis.

关 键 词：枯草芽孢杆菌 绿色荧光蛋白 表达 异源基因 

分 类 号：Q786[生物学—分子生物学]