中国呼吸道合胞病毒地方株G蛋白在重组痘苗病毒中的表达  

Expression of the G glycoprotein from the respiratory syncytial virus field strain by recombinant vaccinia virus

作　　者：邓洁[1] 孙晓鸥[1] 王之樑[1] 陆柔剑[2] 朱汝南[1] 赵林清[1] 钱渊[1] 

机构地区：[1]首都儿科研究所 [2]中国疾病预防控制中心

出　　处：《中华微生物学和免疫学杂志》2004年第4期284-286,共3页Chinese Journal of Microbiology and Immunology

摘　　要：目的　构建表达中国呼吸道合胞病毒 (RSV)地方株G蛋白基因的重组痘苗病毒 ,以用于RSV感染防治的研究。方法　用基因克隆技术将中国RSV地方株G蛋白基因插入到痘苗病毒载体中 ,与痘苗病毒共感染获得重组病毒。用免疫印迹、ELISA、蚀斑减少试验等方法检测表达产物的免疫原性及生物活性。结果　RSV地方株G蛋白在痘苗病毒中获得较好的表达 ,表达产物的糖基化程度较高 ,且该重组病毒免疫家兔可诱发特异性抗体产生。蚀斑减少试验证明 ,用该表达产物制备的抗血清具有中和病毒的活性。结论　重组病毒表达的中国RSV地方株G蛋白具有较好的抗原性。Objective To obtain G glycoprotein from respiratory syncytial virus (RSV) field strain. Methods RSV gene fragment encoding the G glycoprotein from the virus field strain isolated from child with acute respiratory infection in Beijing was inserted into the vaccinia virus vector and the recombinant vaccinia virus expressing the G protein of RSV was obtained by co-infecting this vector with vaccinia virus field strain in cell culture. Rabbits were immunized with the recombinant vaccinia virus expressing RSV G glycoprotein. Western blot, ELISA and plaque reduction assay were used to evaluate the antigenicity, immunogenicity and the biological function of the expressed protein. Results The RSV G glycoprotein was expressed by the recombinant vaccinia virus, which was determined by Western blot with anti-G protein specific monoclonal antibody. The high titer antibodies against G protein and purified RSV were detected from the sera collected from immunized rabbits. And the sera from the immunized rabbits can neutralize the plaque formation of the native RSV. Conclusion The data suggested that the expressed G glycoprotein is similar antigenically and immunogenically with those from native RSV.

关 键 词：呼吸道合胞病毒 G蛋白 重组痘苗病毒 基因克隆技术 特异性抗体 

分 类 号：R392[医药卫生—免疫学]