重组SARS-CoV刺突蛋白基因片段的原核表达及纯化  

Over-expression and purification of recombinant spike protein fragment of SARS coronavirus

作　　者：蒲丹 宋文鑫[1] 闫歌[1] 张秉强[1] 唐霓[1] 王丕龙[1] 郑建[1] 黄爱龙[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所,重庆400010

出　　处：《重庆医科大学学报》2004年第4期409-412,共4页Journal of Chongqing Medical University

基　　金：国家"8 63"高科技发展计划项目资助 (编号 2 0 0 2AA2 15 0 15 ) ;重庆市科委项目资助 (编号 7673 )

摘　　要：目的 :冠状病毒 (SARS -CoV)的刺突蛋白 (spikeglycoprotein ,S蛋白 )是病毒的主要结构蛋白之一 ,也是重要的病毒抗原 ,重组S蛋白的表达可用于检测病毒感染后血清抗体。方法 :根据抗原性预测分析结果 ,将S蛋白中抗原决定簇的编码基因重新拼接成两个新的基因 ,通过全基因合成方式直接合成至原核表达载体pET32a(+) ,转化大肠杆菌BL2 1(DE3) ,以IPTG诱导表达 ,Ni-NTA试剂盒纯化目的蛋白。结果 :SDS -PAGE证实重组S蛋白的表达 ,并被纯化。结论 :重组刺突蛋白的表达及纯化 ,可做为检测SARS -CoV抗体的抗原 ,有助于进一步开展SARSObjective:To obtain the spike glycoprotein(S protein) of SARS coronavirus (SARS-Cov) for detection of the corresponding antibody and preparation of diagnostic kits.Methods:Genes encoding S protein were picked out to form a new gene and divided into two parts according to the prediction of antigenicty.They were obtained by nucleotide synthesis and were cloned into expression vector pET32a(+) respectively.Proteins then were induced by IPTG and purified through Ni-NTA.Results:Target proteins were successfully expressed and purified.Conclusion:The recombinant S proteins of SARS-CoV could be of great help in diagnosing the disease and investigating the virus further.

关 键 词：SARS-COV 刺突蛋白 表达 纯化 

分 类 号：R563[医药卫生—呼吸系统]