小鹅瘟病毒VP3基因的原核表达

Expression of VP3 gene of goose parvovirus in Escherichia coli and preliminary analysis of reactogenicity

根据GenBank已发表的小鹅瘟病毒B株基因序列,设计并合成1对特异性引物,经PCR扩增得到了1 457bp的目的片段,并将该片段克隆至pMD-18Tsimple载体中,再将其亚克隆至原核表达载体pGEX-4T-1中,构建重组表达质粒pGEX-VP3。经0.1mmol/L IPTG诱导,目的蛋白在大肠杆菌中得到表达。Western-blot分析表明,该蛋白具有良好的反应原性。为开发相应的基因工程疫苗奠定了理论基础。

According to GenBank published gosling plague virus strain B gene sequence,specific primers were designed for cloning a 1 457 bp gene fragment.The fragment was cloned to the vector pMD-18T simple,then subcloned to prokaryotic expression vector pGEX-4T-1,and constructed a prokaryotic expression plasmid pGEX-VP3.The target protein was expressed in E.coli and induced by 0.1 mmol/L IPTG.The Western-blot analysis showed that the protein has a good reactivity.This is relevant for the development of genetically engineered vaccine and has laid a solid foundation.