猪Ⅱ型圆环病毒Cap基因重组腺病毒表达载体的构建

Construction and identification of recombinant adenovirus containing the cap gene of type Ⅱ porcine circovirus

通过细菌内同源重组的方法成功构建了含有猪Ⅱ型圆环病毒(PCV2)衣壳蛋白Cap基因的重组腺病毒表达载体。首先将Cap基因亚克隆连接到腺病毒穿梭载体pAdTrack-CMV上,再将重组质粒pAdTrack-CMV／Cap用Pme Ⅰ线性化后电转化携带有腺病毒骨架载体pAdeasy-1的大肠杆菌BJ5183感受态细胞,经细菌内同源重组产生重组腺病毒质粒pAdEasy-Cap。为进一步研究PCV2腺病毒活载体疫苗奠定了基础。

Recombinant replication-defective human adenovirus serotype 5 vector containing the Cap gene of type Ⅱ porcine circovirus （PCV2） was constructed using the method of homologous recombination in bacteria. Cap gene was subcloned into adenovirus shuttle vectors pAdTrack-CMV. The positive recombinant plasmids pAdTrack-CMV/Cap were linearized by Pine Ⅰ and electroporated into Ad-BJ5183 competent cells that had already been transformed with adenovirus skeletal vectors pAdeasy-1,the recombinant adenovirus plasmids pAdEasy-Cap were successfully obtained. These results indicated that recombinant adenovirus could be used on animals for PCV2 recombinant adenovirus vaccine.