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.
出处
《中国畜牧兽医》
CAS
北大核心
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]
