G1型A组轮状病毒地方株VP7基因在杆状病毒系统中的表达  

EXPRESSION OF TYPE G1 VP7 FROM A GROUP A ROTAVIRUS FIELD STRAIN IN BACULOVIRUS SYSTEM

作　　者：何湘君[1,2] 钱渊[1,2] 靖宇[1,2] 李国华 

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 [2]北京医科大学人民医院中心实验室

出　　处：《病毒学报》1999年第1期75-77,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，每年因轮状病毒腹泻造成的死亡超过１００万人。ＶＰ７是病毒外壳上的主要糖蛋白，具有中和抗原活性，与病毒的毒力及免疫保护性有关。也是划分血清型的主要标志，而对疫苗的研究证明，不同血清型之间的交叉保护作用甚...Reverse transcription and polymerase chain reaction(RT PCR) were used to amplify the coding region of VP7 of a G1 type group A rotavirus field strain CR188 circulated in Beijing. The amplified cDNA was cloned into the downstream of the polyhedrin promoter of transfer Vector pBacPAK8. Sequence analysis showed 95.7% homology of the deduced amino acid between VP7 of CR188 and that of G1 prototype Wa. Insect cells were cotransfected with plasmid pBacPAK8 CR188 and Bsu36I digested BacPAK6 viral DNA to produce recombinant baculoviruses by homologous recombination. Recombinant viruses containing CR188 VP7 gene were identified by PCR using the forward primer on the polyhedrin promoter and the downstream primer of VP7. Specific VP7s with four different molecular weights were detected by Western blot using hyperimmune serum against Wa when the VP7 of CR188 was expressed in insect cells.

关 键 词：轮状病毒 VP7 杆状病毒 蛋白表达 

分 类 号：R373.25[医药卫生—病原生物学]