| 福氏志贺菌多重耐药调节蛋白MarA的原核表达 |
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| 引用本文： | 魏小娟,张继瑜,王松泰,牛建荣,李剑勇,周旭正,吴培星,李金善.福氏志贺菌多重耐药调节蛋白MarA的原核表达[J].畜牧与兽医,2008,40(6):12-15. |
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| 作者姓名： | 魏小娟 张继瑜 王松泰 牛建荣 李剑勇 周旭正 吴培星 李金善 |
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| 作者单位： | 中国农业科学院兰州畜牧与兽药研究所,甘肃,兰州,730050 |
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| 基金项目： | 国家自然科学基金资助项目(30671582). |
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| 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。
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| 关 键 词： | 福氏志贺菌 多重耐药调节蛋白 原核表达 蛋白纯化 |
| 文章编号： | 0529-5130(2008)06-0012-04 |
| 修稿时间： | 2007年5月24日 |
Prokaryotic expression of multidrug-resistance regulatory protein MarA of Shigella flexneri |
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| WEI Xiao-juan,ZHANG Ji-yu,WANG Song-tai,NIU Jian-rong,LI Jian-yong,ZHOU Xu-zheng,WU Pei-xing,LI Jin-shan.Prokaryotic expression of multidrug-resistance regulatory protein MarA of Shigella flexneri[J].Animal Husbandry & Veterinary Medicine,2008,40(6):12-15. |
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| Authors: | WEI Xiao-juan ZHANG Ji-yu WANG Song-tai NIU Jian-rong LI Jian-yong ZHOU Xu-zheng WU Pei-xing LI Jin-shan |
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| Abstract: | To obtain multidrug-resistance regulatory protein MarA of Shigella flexneri,the marA gene was amplified by PCR and subcloned into the vector pET-30a(+).The results of digestion with BamHⅠ and SalⅠ,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. |
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| Keywords: | Shigella fleneri multidrug-resistance regulatory protein prokaryotic expression purification |
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