伪狂犬病毒基因转移载体的快速构建及其瞬时表达

以含有伪狂犬病毒（ｐｓｅｕｄｏｒａｂｉｅｓ ｖｉｕｓ,ＰＲＶ）蛋白激酶（ｐｒｏｔｅｉｎ ｋｉｎａｓｅ,ＰＫ）基因的重组质粒为基础,利用非互补粘性经过部分补平后成为粘性末端,再进行连接的方法克隆了与载体分子大小相当的报告基因表达盒。用限制发现人切酶分析确证了伪狂现毒表达载体中报宽大圊窝囊斩主方向,并通过导入细胞后检测外源基因在体外的瞬时表达,直接鉴定一表达载体的生物学活性。

Rapid Construction of the Gene Transfer Vector of Pseudorabies Virus and Its Transient Expression in vitro

A gene expression cassette with molecular size of the insert, a recombinant plasmid containing the PK gene of PRV,equal to vector was cloned through partial repair of the terminal ends of insert and vector, thus transforming them from nonadherent to adherent ones. The constructed gene expression vector of PRV with the report gene was further identified by endonuclear enzyme analysis and the biological activity of expression vector was recognized directly through transfecting cells to detect its transient expression in vitro.