【摘要】 目的:表达SARS冠状病毒S蛋白片段,并制备特异性单克隆抗体(mAb)。方法:原核表达SARS冠状病毒S蛋白片段,从SARS病毒cDNA中扩增S蛋白(N端205至419aa)基因片段,测序结果正确。将该基因片段插入pQE-30中,构建重组质粒pQE-30-S1m,以重组质粒转化E.coliM15诱导表达His-S1m蛋白。纯化后免疫BALB/c小鼠,经融合、筛选制备特异性mAb。结果:表达并纯化了重组蛋白His-S1m,获得1株抗His-S蛋白的mAb1H2。Ig亚类鉴定为IgG1a,轻链为κ型。经Western blotting检测,mAb1H2与S蛋白发生特异性反应,而与His蛋白无交叉反应。结论:成功表达了重组S蛋白并制备了特异性mAb,为SARS-CoV的早期诊断及进一步研究S蛋白的功能奠定了基础。更多还原

【Abstract】 Objective To generate the recombinant fragment at N-terminal residues 205 to 419aa of SARS coronavirus(SARS-CoV) Spike(S) protein,and the monoclonal antibody specific for S protein.Methods The S gene fragment of SARS-CoV was constructed by PCR techniques and expressed in E.coli.Furthermore,purified recombinant protein was used to immune BALB/c mice.The resulting monoclonal antibody was screened from hybridomas,followed by identification of specificity in ELISA and Western blotting.Results Recombinant S1m protein of SARS-CoV was constructed,expressed and purified.One hybridomas producing mAb specific to S1m protein was generated successfully.It was IgG1a(κ).Conclusion One specific mAb against S1m protein of SARS-CoV has been established,which can be used in S protein research and the early diagnosis of SARS-CoV infection.更多还原