伪狂犬病毒Sa株gI-/gE-/YFP+基因缺失载体构建及突变株筛选

根据伪狂犬病病毒SA株的gI和gE基因序列,设计两对引物,缺失掉gE基因5’端738bp,在克隆测序的基础上,采用酶切方法构建了含部分gE基因的转移载体pBgIE,同时将pBLYFP载体上含CMV启动子、多克隆位点、黄色荧光蛋白(YFP)基因和polyA尾巴的表达盒双酶切后插入到缺失位置,构建转移载体,命名为pBgIE-YFP,为下一步开发以伪狂犬病病毒为载体的基因工程疫苗提供了基础。

Construction of a PRV Sa strain gI-/gE-/YFP+gene deleted transfer vector and mutant screening

According to the sequence of gE and gI gene of pseudurabie virus Sa strain, two pairs of primers were designed, with 738bp deleted in gE gene 5’terminal. On the basis of cloning and sequencing, the transfer plasmid vector pBgIE was constucted to get the gE gene partial deletion mutant. The cassette including CMV promter, MCS, GFP gene and poly A from pBLYFP was inserted into the deleting site. The recombinant plasmid pBgIE-YFP deleted part of gE gene was constructed.It provided the basis for developing gene engineering vaccine based on the recombinant PRV expressing foreign genes.