›› 2010, Vol. 37 ›› Issue (1): 45-50.

• 生物技术 • 上一篇    下一篇

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

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   

  1. （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）
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-01-20 发布日期:2010-01-20
  • 通讯作者: 景志忠

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

JIA Huai-jie1,2, JING Zhi-zhong1, FANG Yong-xiang1, WANG Xiao-xia1, CHEN Guo-hua1, LI Ke-bin1,2, DOU Yong-xi1, QU Hui-ling1,2,3   

  1. (1.State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, CAAS; Key Laboratory of Veterinary Public Health of Ministry of Agriculture, Lanzhou 730046, China; 2.Veterinary College of Gansu Agricultural University, Lanzhou 730070, China; 3.Veterinary Bureau of Gansu Province, Lanzhou 730030, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-01-20 Published:2010-01-20
  • Contact: JING Zhi-zhong

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）。

关键词: 猪圆环病毒2型; 基因组; 克隆; 序列分析

Abstract: 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.

Key words: porcine circovirus type 2(PCV2); genome; cloning; sequence analysis

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