水泡性口炎病毒N基因的克隆及其PCR检测方法的建立

Cloning of N gene of vesicular stomatitis virus (VSV) and establishment of the PCR method for dectection of VSV

根据国外发表的水泡性口炎病毒 (VSV)基因组核苷酸序列 ,设计了 2对特异性引物 ,通过RT PCR方法得到N基因。将此基因与 pGEM TEasyVector相连 ,通过蓝白斑筛选出阳性克隆 ,碱裂解法小剂量提取质粒。经测定 ,N基因与参考序列同源性高达 99%。同时根据克隆产物进行亚克隆 ,建立了VSV的PCR检测方法。

A pair of primers was designed to amplify N gene of VSV according to the published relevant sequence of the virus. The N gene was obtained by RT-PCR and sequenced after being directly inserted into the pGEM-T Easy Vector. The result showed that the homology between N gene and the published sequence was 99%.Based on the cloning of N gene,another pair of primers was designed. Through subcloning,a new PCR method had been established to detect the VSV.