人轮状病毒G2和G3型主要中和抗原VP7基因在重组腺病毒中的表达  

Expression of main neutralization antigen VP7 of human rotavirus serotype G2 and G3 by recombinant adenoviruses

作　　者：魏强[1,2,3] 姜秀丽[1] 王健伟[1] 洪涛[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所腹泻病毒室,院士实验室,北京100052 [2]吉林大学基础医学院与中国疾病预防控制中心病毒病预防控制所 [3]洪涛院士实验室

出　　处：《中华实验和临床病毒学杂志》2004年第1期12-15,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家 8 63计划现代农业与生物技术领域资助项目(2 0 0 1AA2 15 0 11)

摘　　要：目的　研究我国G2和G3型轮状病毒主要中和抗原VP7基因在重组腺病毒中的表达。方法　在前期成功表达G1型VP7的基础上 ,选用我国G2和G3型主要流行株 97S4 3和 97S4 8VP7基因 ,用非复制型腺病毒载体对上述基因进行表达。结果　获得了表达我国G2型和G3型人轮状病毒流行株VP7基因的非复制型重组腺病毒rvAdG2VP7和rvAdG3VP7,应用PCR及Southernblot技术证实在重组腺病毒中整合有轮状病毒G2型VP7和G3型VP7基因 ,RT PCR证明重组腺病毒在感染的 2 93细胞内均能有效地转录插入基因 ,Westernblot检测到轮状病毒VP7基因的表达。结论　这一工作为进一步进行动物实验 ,发展多价轮状病毒疫苗打下了基础。Objective To express main neutralization antigen VP7 of human rotavirus serotype G2 and G3 by recombinant adenoviruses on the basis of previous investigation of the prevalence of rotavirus in China. Methods On the basis of successfully expression of human rotavirus protein G1VP7 by recombinant adenovirus vector,the authors constructed some potent recombinant adenovirus strains encoding rotavirus G2 VP7 and G3 VP7 genes which belong to the main rotavirus isolates 97S43 and 97S48. Results Replication-defective recombinant adenoviruses expressing human rotavirus serotype G2 and G3 VP7 genes,named as rvAdG2VP7 and rvAdG3VP7 were successfully constructed. VP7 genes integrated into the viral genome were identified by PCR and Southern blot assay,and specific transcription were detected by RT-PCR in the 293 cells infected with recombinant adenoviruses. Expression of rotavirus VP7 proteins was demonstrated by Western blot assay. Conclusion The established recombinant adenoviruses expressing G2 and G3 serotype VP7s laid a significant basis for further animal experiments in the development of multivalent rotavirus vaccines against rotavirus infection.

关 键 词：G2型轮状病毒 G3型轮状病毒 VP7基因 重组腺病毒 基因表达 

分 类 号：R346[医药卫生—基础医学]