猪瘟病毒E2基因抗原结构域A、B、C、D区在大肠杆菌中的表达  

Study on the Expression Antigen Region A、B、C、D Domain E2 Gene of Classical Swine Fever Virus in E.coli

作　　者：张永国[1] 刘湘涛[1] 韩雪清[1] 胡建和[1] 张彦明[1] 谢庆阁[1] 

机构地区：[1]中国农业科学院兰州兽医研究所

出　　处：《畜牧兽医学报》2004年第2期182-185,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重大基础研究发展规划项目(G19990119)

摘　　要：采用PCR技术扩增出猪瘟病毒E2基因抗原结构域A、B、C、D区并克隆于PGEM Teasy载体上,测序结果表明插入的为猪瘟病毒E2基因抗原结构域,切出目的基因,将目的基因克隆到表达质粒pProEX HTb中,获得重组质粒经PCR、酶切和序列分析鉴定E2基因抗原结构域插入的位置、大小和读码框均正确,SDS PAGE检测表明受体菌经IPTG诱导后能表达E2基因抗原结构域蛋白,Westernblot检测表明受体菌诱导表达E2基因抗原结构域蛋白可与猪瘟阳性血清发生特异性反应。The antigen region A?B?C?D domain E2 gene of Classical swine fever virus was amplified by the Polymerase Chain Reaction (nPCR).The amplified fragments was cloned into PGEX- T- Easy vector and then sequenced.It showed that the insert fragment was the antigen region A?B?C?D domain E2 gene of CSFV.We had digested the interested gene and cloned it into the digested expressed vector pProEX- HTb.It was identified that the position of the antigen region A?B?C?D domain E2 gene of CSFV insert,the size and the reading frame are all right by PCR,restriction digestion and the sequence analysis.SDS- PAGE indicated that the reciepient germs transducted and induced by the recombinant plasmids could express antigen region A?B?C?D domain E2 gene of CSFV.Western blot indicated that the expressed antigen protein could be recognized by the positive serum of CSFV.

关 键 词：猪瘟病毒 E2基因抗原结构域 A区 B区 C区 D区 大肠杆菌 基因表达 

分 类 号：S852.651[农业科学—基础兽医学]