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猪繁殖与呼吸综合征病毒Hn-1/06株GP3蛋白的原核表达与纯化

DOI: 10.7668/hbnxb.2009.01.023, PP. 103-107

Keywords: PRRSV,GP3蛋白,原核表达

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Abstract:

根据GenBank中PRRSV美洲株ATCCVR2332的ORF3基因序列,利用Primer5.0软件设计合成了1对特异性引物,经RT-PCR从河南分离株Hn-1/06株扩增得到了大小为765bp的片段,并将扩增的片段插入pTG19-T载体进行测序而获得含有ORF3的阳性克隆pTG19-T-GP3。将此基因亚克隆到原核表达载体pET-32a,经筛选获得了阳性重组质粒pET-GP3,进而在IPTG的诱导下成功表达,获得了大小约为45kDa的融合蛋白GP3-His,纯化后经Westernblot检测证实表达的融合蛋白具有良好的生物学活性。从而为进一步研究PRRSV次要结构蛋白GP3的免疫特性和功能奠定了基础。

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