贵州地区羊口疮病毒B2L基因克隆及原核表达载体构建  

Cloning and Construction of Prokaryotic Expression Vector of B2L Gene of Orf Virus in Guizhou

作　　者：向智龙[1] 卓建华 程振涛[1,3] 欧德渊[1,3] 鲜思美[1,3] 尹传宝[1] 黄璐[1] 禾彩红[1] 

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]京山县畜牧兽医局,湖北京山431800 [3]贵州省动物疫病研究所,贵州贵阳550025

出　　处：《中国畜牧兽医》2011年第7期76-79,共4页China Animal Husbandry & Veterinary Medicine

基　　金：贵州省科学技术基金项目“贵州省羊口疮的病原学与检测技术研究”(黔科合J字[2010]2260);贵州大学引进人才科研项目“贵州省羊口疮的诊断与防控技术研究”(贵大人基合字[2009]024号);贵州大学2010年研究生创新基金项目“贵州省羊口疮的分子诊断技术及B2L基因表达研究”(校研农[2011]024);贵州大学2010年“SRT”项目“羊传染性脓疱病毒贵州株的PCR鉴定及分子特征分析”[贵大SRT字(2010)021]

摘　　要：用H2L基因特异性引物对羊口疮临床疑似病料进行鉴定,采用PCR扩增出阳性样品羊口疮病毒结构蛋白B2L基因,回收纯化PCR产物,克隆至pMD18-T,测序结果表明插入的片段为目的基因,全长1137 bp。将目的基因定向克隆至pET-32a构建了B2L基因原核表达载体pET-32a-B2L,经PCR鉴定、酶切及进一步测序证明表达载体构建成功,为下一步的表达及基因工程疫苗的研究奠定了良好基础。To identify suspected goats orf case and construction of prokaryotic expression vector of B2L gene of orf virus（ORFV）,clinical and suspect samples of orf could be amplified by a pair of specific primers,then the structural protein B2L gene of orf virus was amplified by PCR method using specific primers.The B2L gene was ligated with pMD18-T vector and the sequencing results showed that the full-length of the inserted gene fragment was 1137 kb.Cutting the target fragment,which was cloned into pET-32a vector and construction of prokaryotic expression vector pET-32a-B2L,the recombinant plasimid was identified by restriction enzyme and PCR,sequence analysis indicated that to be successful construction of pET-32a-B2L expression vector and it has laid a good foundation for the expression and genetic engineering vaccine research.

关 键 词：羊口疮病毒 克隆 原核表达载体 

分 类 号：Q78[生物学—分子生物学]