狂犬病病毒SRV_9株磷酸化蛋白P基因和基质蛋白M基因重组质粒的构建  

Construction of Recombinant Plasmid Phosphorylated Protein Gene P and Matrix Protein M Gene of Rabies Virus SRV_9

作　　者：苏晓慧[1] 夏婷婷[1] 翟少华[1] 简子健[1] 

机构地区：[1]新疆农业大学,乌鲁木齐830052

出　　处：《新疆农业科学》2014年第12期2296-2300,共5页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金(31360623);新疆维吾尔自治区普通高校重点学科(基础兽医学)科研启动基金

摘　　要：【目的】重组狂犬病病毒SRV9株磷酸化蛋白的P基因和基质蛋白的M基因,为构建狂犬病毒(SRV9)全长c DNA提供技术支撑。【方法】设计重组PCR引物,分别扩增SRV9的P基因和M基因,构建重组质粒p MD18-PM,并对重组质粒进行酶切、测序鉴定。【结果】PCR扩增的SRV9的P基因和M基因大小分别为893 bp和608 bp,与Gen Bank已发表的狂犬病病毒SRV9(登录号:AF499686)序列比较,同源性分别为99.66%和99.67%。融合基因PM片段大小为1 501 bp,与预期大小相同,且融合完全。【结论】应用重组PCR成功融合狂犬病病毒SRV9磷酸化蛋白P基因和基质蛋白M基因,为后续构建病毒全长c DNA及感染性克隆奠定基础。[ Objective ] The recombinant genes encoding phosphorylated protein （P）and matrix protein （M） of rabies virus SRV9 were constructed in the paper for building full - length eDNA of this virus. [ Method] The phosphoprotein gene P and matrix protein gene M of rabies virus SRV9 were amplified by the recombinant PCR to construct recombinant plasmid pMD18 - PM. The recombinant product was identified by enzymatic digestion and sequencing. [ Result ] Amplified P and M genes of SRV9 were 893 bp and 608 bp, and homology was 99.66% and 99.67%, respectively, compared with the SRV9 rabies virus sequenee published in the GenBank （ID： AF499686）. The two genes were completely fused and its size was 1 501 bp as same as expec- ted one. [ Conclusion] The genes P and M of rabies virus SRV9 were successfully eonfused to develop a simpler,faster, more applicable method for constructing the full -length eDNA of rabies virus SRV9.

关 键 词：狂犬病病毒SRV9 磷酸化蛋白 基质蛋白 重组PCR 基因融合 

分 类 号：S852.65[农业科学—基础兽医学]