SARS-CoV S1蛋白基因克隆及其在Vero E6细胞中的表达  

Clone and expression of SARS-CoV S1 gene in Vero E6 cells

作　　者：黎万玲[1] 倪兵[1] 李晋涛[1] 李艳秋[2] 王希良[2] 姜曼[1] 肖宇[1] 吴玉章[1] 

机构地区：[1]第三军医大学免疫学教研室,重庆400038 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《免疫学杂志》2003年第4期247-249,257,共4页Immunological Journal

摘　　要：目的　克隆表达SARS CoVS1蛋白 ,构建SARS基因疫苗。方法　从SARS病毒的cDNA中以PCR方法扩增编码人SARS CoVS1蛋白的编码序列 ,将该序列克隆入真核表达载体pVAX1,构建重组表达载体。采用脂质体法转染VeroE6细胞 ,用Westernblot检测S1蛋白的表达。结果　PCR方法扩增出 2 0 0 0bp左右的基因片段 ,插入pVAX1构建重组表达载体后 ,经序列测定证实扩增的片段为SARS CoVTor 2株S1蛋白编码序列。将重组子转染VeroE6细胞 ,收集细胞总蛋白 ,Western检测获得特异蛋白带。结论　成功克隆并表达了SARS CoVS1蛋白 。Objective To clone and express SARS-CoV S1 protein and to construct SARS gene vaccine.Methods SARS-CoV S1 gene was amplified from the cDNA of SARS-CoV by PCR and the gene fragment was cloned into vector pVAX1 to construct recombinant expressing plasmids. The recombinant was transferred into Vero E6 cells by DOTAP reagent, then the expression of S1 protein was detected with Western blot.Results The gene fragment of about 2 kb was produced by PCR. Following cloned into vector pVAX1, sequencing result indica-ted that the sequence was identical to that of S1 derived from Tor 2 strain. After the recombinant was transformed into Vero E6 cells, Western-blot showed specific protein band with the anticipated molecular weight.Conclusion SARS-CoV S1 gene has been cloned and expressed successfully which has been the groundwork for the study of SARS gene vaccine.

关 键 词：SARS-CoVS1蛋白 基因克隆 VeroE6细胞 表达 

分 类 号：R392.1[医药卫生—免疫学]