猪肺炎霉形体Dnak-P97融合基因的表达及其产物的生物学活性  

Expression of Dnak-P97 fusion gene of Mycoplasma hyopneumoniae and biological activities of the expressed product

作　　者：王亮[1] 乔祖健[1] 李媛[1] 张建华[1] 梁立[1] 辛九庆[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001

出　　处：《中国兽医科学》2008年第2期123-127,共5页Chinese Veterinary Science

基　　金：农业部财政专项

摘　　要：应用DNA重组技术,将猪肺炎霉形体黏附因子P97 C末端基因与猪肺炎霉形体伴侣蛋白DnakC末端基因同时克隆到pET-30a表达载体上,构建了重组表达载体。将鉴定正确的重组质粒转化大肠杆菌BL21,用终浓度为1.0 mmol/L的IPTG诱导6h。用BugBusterTMNi-NTA His.Bind纯化试剂盒在自然条件下对目的蛋白进行了纯化;经Western-blotting和动物试验,证实,该蛋白具有良好的生物学活性。The C-terminal Mof P97 gene and C-terminal Dnak gene of Mycoplasma hyopneumoniae （Mhp） were linked and co-cloned into pET-30a expression vector by DNA recombination technique to construct recombinant vector P97-Dnak-pET-3a. The positive recombinant plasmids were transformed into host strain Escherichia coli BL21 and induced with 1.0 mmol/L IPTG for 6 hours. The expressed protein was purified with BugBuster^TM Ni-NTA His · Bind purification kit and its biological activities were identified by Western-blotting and experimental infection of animals.

关 键 词：猪肺炎霉形体 猪霉形体肺炎 P97基因 伴侣蛋白Dnak 热休克蛋白70 

分 类 号：S852.62[农业科学—基础兽医学] Q786[农业科学—兽医学]