猪胸膜肺炎放线杆菌外膜脂蛋白基因的原核表达

Prokaryotic expression of Actinobacillus pleuropneumoniae out membrane lipoprotein gene

经PCR从含有OML基因片段的重组质粒App-OML-1中扩增出mAppOML-1基因,同时对目的基因和表达载体pPRoEX HTb进行酶切、连接、转入E.coliBL21,构建重组质粒pPRoAppOML-1.阳性克隆用IPTG进行诱导,经SDS-PAGE电泳鉴定,表达的融合蛋白分子质量约为45 ku,Western-blotting分析表明表达的目的蛋白具有良好的反应原性.

The mAppOML-1 gene was amplified from the recombinant plasmid App-OML-1 by PCR. The target gene and expression vector pPRoEX HT were digested and ligated,then transformed into E. coli BL21 for constructing the recombinant plasmid pPRoAppOML-1. Positive clones were induced by IPTG. And 45 ku molecular weight of expressed fusional protein was detected by SDS-PAGE. Results showed that the target protein has good reactiongenicity.