福氏志贺菌多重耐药调节蛋白MarA的原核表达  

Prokaryotic expression of multidrug-resistance regulatory protein MarA of Shigella flexneri

作　　者：魏小娟[1] 张继瑜[1] 王松泰[1] 牛建荣[1] 李剑勇[1] 周旭正[1] 吴培星[1] 李金善[1] 

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所,甘肃兰州730050

出　　处：《畜牧与兽医》2008年第6期12-15,共4页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金资助项目(30671582)

摘　　要：为获得福氏志贺菌多种耐药调节蛋白MarA,将扩增出的marA基因亚克隆到原核表达载体pET-30a(+)中,经PCR和双酶切鉴定以及序列测定,重组质粒构建成功。将重组质粒转化到大肠杆菌BL21(DE3)中进行表达,表达的蛋白再经Ni-NTA亲和层析纯化,结果显示,MarA蛋白的表达量占菌体总蛋白的30.2%,经Western-bloting检测,表达的蛋白能被福氏志贺菌阳性血清特异识别,说明表达蛋白具有良好的免疫反应性。To obtain multidrug-resistanee regulatory protein MarA of Shigellaflexneri, the marA gene was amplified by PCR and subeloned into the vector pET-30a （ ＋ ）. The results of digestion with BamH I and Sal I , PCR identification and sequencing analysis showed that the recombinant plasmid was successfully constructed. Then the recombinant plasmid was transformed into E. coli L21 （ DE3 ） competent cells for expression. The expressed protein was purified with Ni-NTA Column under denaturing conditions. The expressed MarA accounted for 30.2% of the total bacterial proteins. Western-blotting revealed that the expressed protein could be recognized specifically by antiserum against S.flexneri, which suggested the expressed protein MarA had a good immunoreactivity.

关 键 词：福氏志贺菌 多重耐药调节蛋白 原核表达 蛋白纯化 

分 类 号：S852.61[农业科学—基础兽医学]