小反刍兽疫病毒H、F基因及其植物表达载体相关序列克隆

Cloning of Peste des petits ruminants virus gene H,F and their related gene sequences in plant expression vectors

为了构建小反刍兽疫病毒(PPRV)植物表达载体,试验采用TA克隆的方法,从PPRV质粒中分别扩增出F、H基因,在H基因终止密码子前加入KDEL序列,形成H-KDEL,从烟草SRⅠ中扩增出促进外源基因表达的MAR12序列和MAR34序列,从pHL005质粒中扩增出报告基因——绿色荧光蛋白GFP基因,以上克隆载体均带有相应酶切位点。结果表明:所克隆的基因和序列大小均与理论值相符,说明各克隆载体均构建成功。

To construct plant expression vectors of Peste des petits ruminants virus （PPRV）,the genes of F and H were amplified from PPRV plasmid,H-KDEL through adding KDEL was formed into the gene H before its termination codon,two sequences MAR12 and MAR34 in Tobacco SRⅠwhich enhance the expression of exogenous genes,as well as a reporter gene,green fluorescent protein gene from pHL005 plasmid was cloned by TA cloning.All the clones were with the enzyme site.The result showed that the clones size of all the genes and sequences were the same to the theories.The cloning vectors had been successfully constructed.