大肠杆菌F18菌毛FedE蛋白的表达与鉴定  

Expression and identification of the minor subunit FedE of the fimbriae F18 of Escherichia coli

作　　者：成大荣[1] 徐建生[1] 孙怀昌[1] 高崧[1] 刘俊斌[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2005年第1期5-8,共4页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家高技术发展计划项目(863-2003AA222141)

摘　　要：根据已经发表的F18菌毛E亚单位的基因序列(fedE)设计1对引物,利用PCR技术从重组质粒TF107E中扩增到一段序列,并按预定的阅读框架插入到表达性质粒载体pGEX-6p-1中的谷胱苷肽转移酶(GST)基因的下游,获得重组质粒pPFedE,并转化大肠杆菌BL21获得重组菌PPFedE。琼脂糖凝胶电泳、序列测定及分析表明:该序列大小为459bp,与GENEBANK中的FedE结构编码序列(Z26520)完全一致。通过对菌体裂解物的SDS-PAGE电泳分析以及Westernblotting鉴定,证明重组大肠杆菌PPFedE的可以表达融合蛋白形式的FedE(命名为GST-FedE),即FedE蛋白(16.297ku)与谷胱苷肽转移酶(27.335ku)相连组成分子量为43.632ku的融合蛋白。A pair of primers was designed and synthesized according to the minor subunit FedE gene of fimbriae F18, and the subgenic fragment of fedE was amplificated from the recombined plasmid TF107E. with appropriate restriction sites. The plasmid vector pGEX-6p-l was used for the expression of the fedE. The 5' terminus of the gene that code for fedE was genetically fused to the 3' terminus of the gene coding for the enzyme glutathione S-transferase , which serves as a carrier in this expression system. The resulting plasmid contained the fedE gene was designed as pPFedE. The pPFedE was transferred into E. coli BL21 and confirmed that it was able to express large quantities of a 43. 632 ku fusion protein (GST-FedE) by SDS-PAGE analysis and Western-blotting.

关 键 词：大肠杆菌 菌毛 F18 FedE 表达 鉴定 

分 类 号：S855.1[农业科学—临床兽医学] Q786[农业科学—兽医学]