【摘要】 目的克隆分析中国特有小型猪来源的猪内源性反转录病毒(PERV)囊膜基因(env),比较不同来源毒株的异同及系统进化关系。方法应用RT-PCR的方法分别从中国实验小型猪、巴马香猪及五指山小型猪外周血淋巴细胞中扩增PERVenv基因,克隆入T载体后测序;随后以PCR方法对克隆PERVenv基因进行分型检测;最后采用DNAsis及DNAstar软件对克隆的env基因和国外毒株env基因进行同源性及系统进化进行分析比较。结果本试验所克隆的3株PERVenv基因长度分别为1983bp、1986bp及1968bp,分别编码661、662和656个氨基酸,分型检测均为PERV-A亚型。同源性分析表明:国内这3个分离株env基因与国外来源于不同宿主的PERV-A、B、C亚型env基因的同源性分别在92.8~96.5%、69.4~73.4%及77.5%~80.8%之间。结论PERV中国株的进化与A、B、C亚型分类有关,与宿主、分布地域关系不大,且PERV-A亚型与C亚型的亲源关系更近于B亚型。更多还原

【Abstract】 To analyze the interrelation and variation between the envelope gene of porcine endogenous retrovirus from miniature swine of China and other PERV published strains.Total RNA was extracted from peripheral blood lymphocyte of miniature swine by TRIzol reagent.The envelope gene was amplified by RT-PCR,then the amplified fragments were cloned into pGEM-T and sequenced subsequently.The result showed that three copies of PERV env gene with the length of 1983bp,1986bp and 1968bp,encoding 661,662 and 656 amino acides respectively,were identified to be PERV-A subtype.The nucleotide sequence homology between this Chinese strain and the published strain class A,B and C was 92.8-96.5%,69.4-73.4% and 77.5%-80.8% respectively.It is concluded that Phylogenetic Analysis revealed that the evolution of PERV Chinese strain was related to three subtypes of PERV but there was little relationship to host and area distribution,and genetically class A is more related to class C than class B.更多还原