SARS冠状病毒S蛋白基因片段的表达及其单克隆抗体的制备  

Generation of recombination N-terminal residues 205 to 419 of SARS coronavirus spike protein and its specific monoclonal antibody

作　　者：鲍海逊[1] 夏梅[1] 缪凤琴[1] 谢维[1] 张建琼[1] 

机构地区：[1]东南大学发育与疾病相关基因教育部重点实验室,遗传与发育生物学系,江苏南京210009

出　　处：《东南大学学报（医学版）》2009年第1期18-21,共4页Journal of Southeast University(Medical Science Edition)

基　　金：江苏省卫生厅SARS专项基金资助项目(H200309)

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。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 Slm protein of SARS-CoV was constructed, expressed and purified. One hybridomas producing mAb specific to Slm protein was generated successfully. It was IgG1 a （K）. Conclusion One specific mAb against S1 m protein of SARS-CoV has been established, which can be used in S protein research and the early diagnosis of SARS-CoV infection.

关 键 词：严重急性呼吸综合征 SARS冠状病毒 S蛋白 单克隆抗体 

分 类 号：R392.11[医药卫生—免疫学]