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.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
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)
