中国特有小型猪内源性反转录病毒囊膜基因的克隆及进化分析  

Cloning and phylogenetic analysis of envelope gene of porcine endogenous retrovirus derived from miniature swine of China

作　　者：吴健敏[1] 吕茂民[1] 陈凤莲[2] 谢放[1] 潘汉世[2] 阳玉彪[1] 马玲[2] 章金刚[1] 

机构地区：[1]军事医学科学院野战输血研究所 [2]广西兽医研究所

出　　处：《中国人兽共患病学报》2006年第10期925-928,共4页Chinese Journal of Zoonoses

基　　金：国家"十五"重点攻关项目(2004BA717B-02-04);国家自然科学基金(30270989);广西壮族自治区应用基础研究专项基金(桂科基0448041)资助项目

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PERV env基因，克隆入T载体后测序；随后以PCR方法对克隆PERV env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PERV env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。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.

关 键 词：猪内源性反转录病毒 囊膜基因 序列分析 

分 类 号：R382.5[医药卫生—医学寄生虫学]