鹅细小病毒SS/10株VP3蛋白的原核表达及鉴定

根据GenBank收录的鹅细小病毒(GPV)基因序列,设计并合成了一对VP3基因扩增引物。采用PCR技术对SS/10株的VP3基因进行扩增,将目的基因和原核表达载体PET-32a分别经HindⅢ和BamHⅠ双酶切后进行连接,获得重组质粒PET-32a-VP3,并将其转化表达菌BL21(DE3)pLysS。经IPTG诱导,SDS-PAGE分析,结果获得了72ku左右的融合蛋白,大部分以包涵体的形式存在于菌体中。Wesern blot结果表明,该蛋白能与GPV阳性血清发生特异性反应,具有良好的反应原性。

Prokaryotic Expression and Identification of VP3 Gene of Goose Parvovirus SS/10 Strain

One pair of primers was designed to amplify VP3 gene according to the sequences of goose parvovirus(GPV)published in GenBank.The PCR product of SS/10 was cloned into the expression vector PET-32a after double enzyme digestion by HindⅢ and BamHⅠ.The recombinant prokaryotic expression plasmid was constructed,and named PET-32a-VP3.The recombinant plasmid was transformed into BL21(DE3) pLysS.The transformed bacteria were induced with IPTG and the expressed protein was analyzed by SDS-PAGE.The fusion protein of 72 ku was expressed.The protein could react with GPV positive serum specifically in a Western blot test,indicating that the expressed protein had good reactogenicity.