牛皮蝇HypoderminA基因在大肠杆菌中的表达  

Expression of hypodermin A gene of H. lineatum in Escherichia coli

作　　者：王文龙[1] 哈斯阿古拉[2] 阮风雷[1] 呼和巴特尔[1] 赵云刚 

机构地区：[1]内蒙古农业大学动物科学与医学学院,内蒙古呼和浩特010018 [2]内蒙古大学生命科学学院,内蒙古呼和浩特010021 [3]呼和浩特市动物疫病防控中心,内蒙古呼和浩特010020

出　　处：《中国兽医学报》2008年第12期1394-1397,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30360082)

摘　　要：从牛皮蝇Ⅱ期幼虫中提取总RNA,进行RT-PCR扩增牛皮蝇Hypodermin A基因。将扩增基因进行克隆测序,构建原核表达载体pET30-HA,转化大肠杆菌BL21(DE3)。用IPTG诱导表达,得到表达量达到全菌蛋白35%以上的特异性蛋白。SDS-PAGE分析表明,表达产物大部分以包涵体形式存在。Western-blot检测结果表明,获得的重组蛋白为重组Hypodermin A,具有较好的免疫活性。Total RNA was extracted from the second stage larva of H. lineatum. Hypodermin A gene of H. lineatum was amplified by RT-PCR. The target gene was cloned and sequenced. The prokaryotic expression vector pET30-HA was constructed and then transformed to E. coli BL21 （DE3）. The recombinant E. coli was induced and expressed by IPTG. The expression level of the specific protein was more than 35% of total protein. It was mainly present in inclusion body observed by SDS-PAGE. Western blot analysis indicated that the obtained protein was recombinant hypodermin A,and had good reactiogenicity against positive serum.

关 键 词：牛皮蝇 HYPODERMIN A 原核表达 WESTERN-BLOT 

分 类 号：Q78[生物学—分子生物学] S852.74[农业科学—基础兽医学]