羊传染性脓疱病毒B2L基因片段重组腺病毒载体的构建

Construction of Recombinant Adenovirus Vector Containing CEVB2L Gene

[目的]利用腺病毒载体系统构建羊传染性脓疱病毒B2L基因重组腺病毒载体。[方法]以从羊传染性脓疱病毒株JLSY04中提取的基因组DNA为模板,PCR扩增获得B2L目的基因片段;然后将B2L目的基因克隆至PDNR-CMV载体,筛选阳性克隆获得质粒CTC572-6;再将质粒CTC572-6与腺病毒载体进行同源重组,筛选阳性克隆,并进行菌液PCR、酶切、测序等鉴定。[结果]经酶切和基因测序等鉴定,成功构建了携带羊传染性脓疱病毒B2L基因的重组腺病毒载体CTC572Ade-30。[结论]为羊传染性脓疱基因工程疫苗的进一步研究奠定基础。

[Objective] Sheep contagious pustular dermatitis virus B2L recombinant adenovirus was built by adenovirus vector system. [Method] Genome DNA extracted from sheep contagious pustular dermatitis virus strain as a template,gene fragments obtained from B2L by PCR amplification; B2L gene was cloned into PDNR-CMD vector,screening positive clones and plasmid CTC572-6 was obtained; CTC572-6 plasmid for homologous was recombined with the adenoviral vector. Screening positive clones and bacilli PCR,digestion and sequencing and so on were identified. [Result] After identified by enzyme digestion and gene sequencing,recombinant adenovirus vector CTC572Ade-30 of carrying sheep contagious pustular dermatitis virus B2L gene was constructed successfully. [Conclusion] The study laid the foundation for sheep contagious pustular dermatitis genetically engineered vaccine.