基于重组工程法高转化效率的大肠杆菌BL21(DE3)表达菌株构建  

Construction of Higher Conversion Efficiency Escherichia coli BL21( DE3) via Recombineering

作　　者：张飞飞[1] 石牡丹[2] 尚广东[2] 

机构地区：[1]徐州医学院江苏省麻醉学重点实验室,江苏徐州221004 [2]江苏省微生物工程技术研究中心,江苏省微生物与功能基因组学重点实验室,南京师范大学生命科学学院,江苏南京210023

出　　处：《安徽农业科学》2015年第20期29-31,72,共4页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金(NSFC81273412)

摘　　要：[目的]提高最常用的异源蛋白表达宿主菌E.coli BL21(DE3)的转化效率。[方法]利用基于质粒的重组工程系统和双链断裂修复功能。[结果]敲除了编码降解外源DNA的I型限制性内切酶的hsdR基因,获得了高转化效率以及其他功能与BL21(DE3)仍保持一致的LS1928菌株。[结论]获得的LS1928菌株可能会作为获得催化用酶的关键材料,进而在蛋白质工程等研究领域有着重要的作用。[Objective] The aim was to improve the conversion efficiency of the most commonly used heterologous protein expression host strain. [Method] Type I restriction endonuclease capable of degrading exogenous DNA coding gene hsdR was knocked out via plasmid-based recombineering system as well as its inherent double strand break repair（ DSBR） function. [Result]The obtained strain LS1928 was still consistent with E. coli BL21（ DE3） in every aspect except higher conversion efficiency. [Conclusion]The newly developed strain has the potential to be used as key material to obtain catalytic enzyme and hence plays an important role in protein engineering.

关 键 词：重组工程 基因敲除 双链断裂修复 大肠杆菌BL21(DE3) 

分 类 号：S188[农业科学—农业基础科学]