【摘要】 猪流感病毒(swine influenza virus,SIV)不仅给养猪业造成重大经济损失,而且威胁人类健康。血凝素(hemag-glutinin,HA)是SIV囊膜上的重要免疫原,介导病毒吸附和膜融合。针对流感病毒的中和抗体主要以HA靶标阻断受体结合,本研究通过RT-PCR扩增了猪流感病毒A/Swine/Inner Mongolia/547/01(H3N2)的HA基因,并将其克隆入pMD18-T载体。重组pMD18-T经KpnI和PstI酶切后得到的HA基因插入pMelBacA载体的KpnI/PstI位点。PCR和限制性内切酶分析鉴定表明成功构建了pMelBacA-HA转移载体。这为开发HA亚单位疫苗奠定了基础。更多还原

【Abstract】 Swine influenza virus(SIV)can cause enormous economic loss to pig industry and poses threat to human health.The hemagglutinin(HA) is a major immunogen on the envelope of SIV and can mediate viral attachment and membrane fusion.HA is the main target of neutralizing antibodies against influenza virus that act primarily by blocking receptor binding.In the study,HA gene of A/ Swine/ Inner Mongolia/ 547/ 01(H3N2) was amplified by RT-PCR.The product of RT-PCR was cloned into pMD18-T vector.HA gene was inserted into the Kpn I/Pst I sites of pMelBacA after the recombinant pMD18-T was cleavaged with restriction endonuclease Kpn I and Pst I.Identification by PCR and restriction-endonuclease analysis showed that the recombinant transfer vector pMelBacA-HA was constructed successfully.It is helpful for development of subunit vaccine based on HA.更多还原