在昆虫细胞中表达G2型轮状病毒地方株VP7基因  

EXPRESSION OF VP7 FROM A GROUP A ROTAVIRUS G2 FIELD STRAIN IN INSECT CELLS

作　　者：何湘君[1] 钱渊[1] 李国华[1] 靖宇[1] 刘军[1] 

机构地区：[1]首都儿科研究所,北京市感染与免疫中心实验室,北京医科大学人民医院中心实验室

出　　处：《病毒学报》1998年第4期374-376,共3页Chinese Journal of Virology

基　　金：国家自然科学基金;卫生部科研基金;北京市自然科学基金

摘　　要：Ａ组轮状病毒是导致婴幼儿重症腹泻的最主要病毒病原，ＶＰ７是病毒外壳上的主要糖蛋白，它具有中和抗原活性，与病毒的毒力及免疫保护性有关，也是划分病毒血清型的最主要标志之一〔１〕。ＶＰ７基因及其编码蛋白一直是人们研究的主要对象，很多研究工作和诊断试剂都需大...The full length of G2 type VP7 cDNA was amplified by reverse transcription and polymerase chain reaction(RT-PCR)from a group A rotavirus field strain CR100 circulating in Beijing. The amplified cDNA was directly cloned into the transfer vector pBacPAK8 downstream of the polyhedrin promoter. Sequence analysis showed 95.7% homology of the deduced amino acid between VP7s of CR100 and prototype strain Hu-5. Insect cells were cotransfected with recombinant plasmid pBacPAK8-CR100 and Bsu36I-digested BacPAK6 viral DNA. Recombinant baculovirus containing CR117 VP7 gene was obtained by plaque screening and PCR identification using the forward primer on the polyhedrin promoter and the downstream primer of VP7. Specific VP7 with three different molecular weights was detected by Western blot using hyperimmune serum against rotavirus strain DS-1(G2 type)when expressed in insect cells.

关 键 词：轮状病毒 VP7 杆状病毒载体 蛋白表达 

分 类 号：R373.2[医药卫生—病原生物学] Q78[医药卫生—基础医学]