轮状病毒VP4、VP7基因的克隆与核苷酸序列分析  

Cloning and Sequencing of Rotavirus VP4、VP7

作　　者：王鹏雁[1] 陈创夫[1] 余兴龙[2] 徐兴然[2] 涂长春[2] 张高轩[1] 

机构地区：[1]新疆石河子大学动物科技学院,新疆石河子832003 [2]解放军军需大学军事兽医研究所,吉林长春130062

出　　处：《中国预防兽医学报》2003年第2期99-103,共5页Chinese Journal of Preventive Veterinary Medicine

摘　　要：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 816bp、916bp的VP4、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_V4、PT_V7中含有轮状病毒的VP4、VP7基因片段。经核苷酸序列分析 ,表明克隆了轮状病毒主要保护性抗原VP4。The antigenic determinants of Rotavirus VP4 and VP7 genes were amplified from cell infected with rotavirus by reverse transcription_polymerase chain reaction(RT_PCR),the length of target genes was 81bp and 91bp.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 fragments of VP4 and VP7 were cloned and the recombinant plasmids PT_V4 and PT_V7 were constructed.The results of nucleotide sequencing showed that the inserted genes had positive reading frame.

关 键 词：核苷酸序列分析 轮状病毒 基因克隆 VP4 VP7 

分 类 号：S852.659.5[农业科学—基础兽医学] Q785[农业科学—兽医学]