H3N2亚型犬流感病毒HA1基因的原核表达及抗原性分析

摘要

为原核表达H3N2亚型犬流感病毒((CIV) HA1蛋白,本研究利用特异性引物扩增CIV H3分离株的HAI基因,将其克隆到pMD 18-T载体后进行序列测定.再将其亚克隆于pET-32a(+)中构建重组表达质粒pET-HA1.将该质粒转化于大肠杆菌BL21(DE3)中,经IPTG诱导,SDS-PAGE电泳分析,表达的重组蛋白约为58ku.纯化的HA1蛋白经westem blot和Dot-ELISA鉴定表明,表达的重组HAI蛋白可以与H3N2亚型CIV阳性血清发生特异性反应.%To express canine influenza virus (CIV) swbtype H3N2 HA1 protein, the HA1 gene was amplified by PCR and cloned into vector pMD18-T and then sequence determination. The HA1 gene was cloned into vector pET-32a (+). The recombinant plasmid pET-HA1 was transformed into BL21 (DE3) and induced with IPTG. The expressed HA1 fusion protein had a MW of 58 ku and was purified. Western blot and Dot-ELISA showed that the purified HA1 protein could be recognized by CIV positive serum.