J亚群鸡白血病病毒AH09/2株的gp85基因的克隆与原核表达  

Cloning and expression of gp85 gene of avian leukosis virus subgroup J AH09/2 strain

作　　者：刘迪[1] 王桂军[1] 赵圆圆[1] 辛伟[1] 魏建忠[1] 李郁[1] 孙裴[1] 张训海[2] 秦爱建[3] 

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]安徽科技学院,安徽凤阳233100 [3]扬州大学兽医学院,江苏扬州225009

出　　处：《中国微生态学杂志》2010年第8期679-681,共3页Chinese Journal of Microecology

基　　金：农业公益性行业科研专项(20080319)资助;安徽省教育厅自然科学基金项目(KJ2007B176)

摘　　要：利用PCR方法扩增出J亚群禽白血病病毒(ALV-J)AH09/2株的gp85基因全长930 bp DNA片段。经T载体克隆测序并连接到pGEX-6p-1载体上,构建了重组表达质粒pGEX-6P-1-gp85,在IPTG的诱导下进行表达。Western-blot结果分析表明,gp85融合蛋白表达产物分子量大小约61 kDa,并能与ALV-Jenv基因单抗发生特异性反应。这些结果为深入研究GP85蛋白的生物学功能及研制ALV-J检测ELISA试剂盒奠定了基础。The DNA fragment encoding gp85 of avian leukosis virus subgroup J AH09/2 strain was amplified by PCR and cloned into pGEX-6P-1 expression plasmid.After sequencing,the recombined expression plasmid was transformed into E.coli BL21 and induced by IPTG.SDS-PAGE analysis showed that the recombinant fusion protein was efficiently expressed with the molecular weight of 61 kDa.Western-Blot results suggested that the expressed protein could react specifically with the ALV-J env monoclonal antibody.These results will lay a foundation for studying the biological characterization of gp85 gene and developing the ALV-J ELISA test kit.

关 键 词：J亚群禽白血病病毒 GP85基因 原核表达 

分 类 号：S855.3[农业科学—临床兽医学]