山羊痘病毒P32基因重组真核表达载体的构建与鉴定  

Construction and Identification of Eukaryotic Expression Vector of Goat Poxvirus P32 Gene

作　　者：殷俊磊[1] 周碧君[1,2] 文明[1,2] 程振涛[1,2] 樊翠华[1] 杨颖[1] 杜海燕[1] 

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病研究室,贵州贵阳550025

出　　处：《河南农业科学》2010年第9期132-134,共3页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金项目(30940054);贵州省优秀科技教育人才省长专项基金项目(黔省专合字[2006]12号)

摘　　要：根据已发表的山羊痘病毒P32基因序列设计合成1对带有BamHⅠ和HindⅢ酶切位点的引物,用PCR方法从分离的贵州山羊痘病毒LD毒株中扩增出含P32基因的DNA片段,纯化后,经BamHⅠ/HindⅢ双酶切,克隆至真核表达载体pcDNA3.1(+),获得pcDNA3.1(+)-P32/LD重组载体。通过PCR、双酶切及测序鉴定,证实重组真核表达载体pcDNA3.1(+)-P32/LD构建成功。A pair of primers containing the restriction sites for BamHⅠand Hind Ⅲ was designed based on the previously reported sequence of the goat poxvirus P32 gene.The P32 gene DNA sequence was amplified by PCR from the Lab-kept bacterial strains.PCR products were double-digested by BamHⅠand Hind Ⅲ,then cloned into the eukaryotic expression vector pcDNA3.1（＋）.After identification by PCR amplification,double restriction digestion and sequencing,the recombinant expression vector pcDNA3.1（＋）-P32/LD was successfully c onstructed.The experiment will provide help for future research in GPV P32 genetic vaccine and diagnostic method.

关 键 词：山羊痘病毒 P32基因 真核表达载体 PCDNA3.1(+) 

分 类 号：S858.27[农业科学—临床兽医学]