猪圆环病毒2型甘肃兰州株全基因组序列的克隆及序列分析  

Cloning and Sequence Analysis of the Complete Genome of Porcine Circovirus Type 2 Gansu Lanzhou Strain

作　　者：贾怀杰[1,2] 景志忠[1] 房永祥[1] 王晓霞[1] 陈国华[1] 李克斌[1,2] 窦永喜[1] 瞿惠玲[1,2,3] 

机构地区：[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部兽医公共卫生重点实验室,兰州730046 [2]甘肃农业大学动物医学院,兰州730070 [3]甘肃省兽医局,兰州730030

出　　处：《中国畜牧兽医》2010年第1期45-50,共6页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然基金项目(30871884);国家支撑计划(2006BAD31B03);甘肃省支撑计划(0804NKCA076)

摘　　要：参照GenBank中登录的猪圆环病毒2型(PCV2)全基因组序列,设计合成了2对特异性引物,从甘肃兰州疑似断奶仔猪多系统衰竭综合征(PMWS)病料中提取PCV2基因组DNA,分段PCR扩增全长基因组。回收PCR产物,将其插入pGEM-T Easy载体,构建了重组质粒pGEM-PCV2,转化后筛选、提取阳性重组质粒进行测序。结果表明,克隆到的PCV2全基因组长为1767 bp。应用DNAStar序列分析软件对所测PCV2序列与GenBank中登录的包括甘肃在内的国内外PCV2毒株进行同源性分析。结果显示,克隆的PCV2与ZhouKou(EU656143)毒株遗传关系较近,其核苷酸和推导的氨基酸序列同源性高达99.4%、98.6%,与已报道的GS03(EU547458)甘肃毒株遗传关系较远,可能是流行于甘肃兰州的一个变异毒株GSLZ(GenBank登录号为:FJ447482)。Two pairs of primers were designed and synthesized according to the full length genome of PCV2 in GenBank.Genomic DNA was extracted from samples of suspected post weaning multisystemic wasting syndrome（PMWS） pig in Lanzhou,Gansu province,and the full length genome of PCV2 was fragmentation cloned by PCR into pGEM-T easy vector,recombinant vector pGEM-PCV2 were constructed and then sequenced.Sequence analysis with DNAStar software showed that the genome of PCV2 Gansu Lanzhou strain is 1767 bp and the result of homology comparison of the full length genome among strains published in GenBank,indicated that it displayed high nucleotide and amino acid identities with ZhouKou（EU656143） strain（99.4% nucleotide identity and 98.6% amino acid identity）,but distinct from GS03（EU547458） which was another Lanzhou epidemic strain.So strain in this test may be a variant strain（GenBank accession No.FJ447482） prevalent in Lanzhou,Gansu.

关 键 词：猪圆环病毒2型 基因组 克隆 序列分析 

分 类 号：Q78[生物学—分子生物学]