口蹄疫病毒全衣壳前体(P1)重组T4噬菌体的构建和鉴定  

Construction and identification of foot-and-mouth disease virus entire capsid protein precursor(P1) gene recombinant T4 bacteriophage

作　　者：田纯见[1] 毕英佐[1] 曹永长[1] 

机构地区：[1]华南农业大学动物科学学院,广东广州510642

出　　处：《中国兽医学报》2007年第5期671-673,678,共4页Chinese Journal of Veterinary Science

基　　金：国家"863"计划资助项目(2002AA245051);广东省重大科技专项资助项目(2004A20403002)

摘　　要：以含完整的口蹄疫病毒全衣壳前体基因(P1)的质粒pP1-T为模板进行PCR扩增获得P1基因片段(约2 200bp),构建整合质粒pR-P1经引物对Pr.78/P1-A2进行PCR扩增筛选插入方向正确的重组质粒,EcoRⅠ酶切和T7启动子测序正确。以pR-P1质粒转化E.coliE2,感染噬菌体φT4-Z1进行同源重组,经溶菌酶依赖性筛选获得整合成功的噬菌体φT4-P1。SDS-PAGE检测重组噬菌体出现约98 000预计大小的条带;Western bloting检测表明,重组噬菌体φT4-P1与口蹄疫阳性血清可发生免疫反应。With the plasmid pP1-T as template, a entire capsid protein precursor （P1） gene （about 2 200 bp） was obtained by PCR amplification, and the integrative plasmid pR-PT constructed, which was afterwards screened with PCR amplification employing a pair of primers Pr 78/P1-A2 to get recombinants having the correct orientation, and confirmed with EcoR Ⅰ- restriction and nucleotide sequencing of the T7 promoter. E. coli E2 transformed with the plasmid pR-P1 was transfected into the bacteriophage φT4-Z1 for homologous recombination, when successful integration was manifested by lysozyme-dependent screening the bacteriophage φT4-P1 yielded as confirmed by the recombinant bacteriophage revealing a 98 000 expected band on SDS-PAGE, and reacting with foot-and-mouth disease positive serum in Western blotting test.

关 键 词：口蹄疫病毒 全衣壳前体基因 T4噬菌体 

分 类 号：S852.65[农业科学—基础兽医学] R535[农业科学—兽医学]