鹅副粘病毒HN蛋白的原核表达与免疫学鉴定

Prokaryotic expression and immunologic identification of hemagglutinin-neuraminidase gene of Goose paramyxovirus E03 strain

以GenBank登录的鹅源副粘病毒GPMV ZJI株基因组全序列为模板,在HN基因开放阅读框上下游设计一对特异性引物,利用RT-PCR方法扩增得到HN基因,将目的基因克隆至pET32a表达载体上,转化至BL21中,通过SDS-PAGE和Western-blot鉴定目的蛋白的表达。由结果可知,通过RT-PCR法扩增出HN基因,大小与预期一致。SDS-PAGE和Western-blot均得到预期条带,说明pET32a表达载体在BL21中成功表达了HN蛋白,为后续相关免疫学研究奠定基础。

The HN gene of goose paramyxovirus（GPMV） E03 strain was amplified by RT-PCR with the primers designed on the basis of HN gene sequence of GPMV ZJI strain,and then was inserted into the prokaryotic vector pET32a for the construction of prokaryotic expression vector,named pET-HN.Then we transformed pET-HN into Escherichia coli BL21,and identified by SDS-PAGE and Western blot.The target gene was amplified by RT-PCR.SDS-PAGE and Western blot showed the target strap of fusion protein.It was apparent that the HN gene was expressed in E.coli at high level;meanwhile,the prokaryotic expression products of this gene would be a good foundation for immunologic identification about GPMV.