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山羊痘病毒P32基因重组真核表达载体的构建与鉴定
引用本文:殷俊磊,周碧君,文明,程振涛,樊翠华,杨颖,杜海燕.山羊痘病毒P32基因重组真核表达载体的构建与鉴定[J].河南农业科学,2010(9).
作者姓名:殷俊磊  周碧君  文明  程振涛  樊翠华  杨颖  杜海燕
作者单位:1. 贵州大学,动物科学学院,贵州,贵阳,550025
2. 贵州大学,动物科学学院,贵州,贵阳,550025;贵州省动物疫病研究室,贵州,贵阳,550025
基金项目:国家自然科学基金项目,贵州省优秀科技教育人才省长专项基金项目 
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。

关 键 词:山羊痘病毒  P32基因  真核表达载体  pcDNA3.1(+)

Construction and Identification of Eukaryotic Expression Vector of Goat Poxvirus P32 Gene
YIN Jun-lei,ZHOU Bi-jun,WEN Ming,CHENG Zhen-tao,FAN Cui-hua,YANG Ying,DU Hai-yan.Construction and Identification of Eukaryotic Expression Vector of Goat Poxvirus P32 Gene[J].Journal of Henan Agricultural Sciences,2010(9).
Authors:YIN Jun-lei  ZHOU Bi-jun  WEN Ming  CHENG Zhen-tao  FAN Cui-hua  YANG Ying  DU Hai-yan
Abstract: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.
Keywords:pcDNA3  1(+)
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