轮状病毒VP7基因的克隆及核苷酸序列分析

Gene Cloning and Sequencing of Rotavirus VP7

用反转录聚合酶链式反应 (RT PCR)技术 ,从轮状病毒感染的细胞中扩增了 916bp的VP7片段中抗原表位区。通过T4DNA连接酶将其直接连接于克隆载体质粒pGEM T上 ,转化至受体菌DH5α中。提取质粒经PCR扩增、酶切鉴定 ,证明重组质粒pT V7中含有轮状病毒的VP7基因片段。经核苷酸序列分析 。

The antigenic determinant of Rotavirus VP7 genes was amplified from cell infected by rotavirus by reverse transcription polymerase chain reaction(RT PCR). The lenth of target gene was 916 bp.The products of RT PCR were ligated with plasmid pGEM T and transformated to E.coli DH5α.By the analysis of restriction endonuclease and PCR ,the fragment of VP7 was cloned and the recombinant plasmid PT V7 was constructed.The result of nucleotid sequencing showed that the inserted gene had positive reading frame.