摘要
利用RT PCR和nestedPCR(nPCR)技术扩增出猪水泡病病毒VP1基因的抗原区 ,将其克隆到表达载体pProEX HTb中 ,获得重组质粒 ,经PCR、酶切和序列分析鉴定表明 ,目的基因插入的位置、大小和读码框均正确。将重组质粒导入BL2 1 (DE3) ,经IPTG诱导表达后SDS PAGE检测表明 ,重组菌能表达猪水泡病病毒VP1抗原区蛋白 ;Westernblot检测表明 ,诱导表达的抗原区蛋白能与猪水泡病阳性血清发生特异性反应。
The antigenic region of VP1 gene of swine vesicular disease virus was amplified by reverse transcription polymerase chain reaction (RT-PCR) and nested polymerase chain reaction (nPCR).After the amplified fragment was cloned into the expression vector pProEX-HTb. The insert position,the size and the reading frame of the insertion were identified by PCR,restriction digestion and sequence analysis of the recombinant plasmids.SDS-PAGE and Western blot indicated that the transformed BL21(DE3) by the recombinant plasmids and induced by IPTG could express the antigen region of VP1 of swine vesicular disease virus, the expressed antigen protein could be recognized by the positive serum of SVDV.
出处
《微生物学报》
CAS
CSCD
北大核心
2003年第3期342-346,共5页
Acta Microbiologica Sinica
基金
国家重大基础研究发展规划项目 (G19990 119)~~
关键词
猪
水泡病病毒
VPl基因
抗原区
原核表达
Swine vesicular disease virus, VP1 gene, Antigen region, Expression
