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猪Ⅱ型圆环病毒豫A株的全基因组克隆与序列分析
引用本文:曹胜波,陈焕春,肖少波,赵俊龙,吕建强,钱平.猪Ⅱ型圆环病毒豫A株的全基因组克隆与序列分析[J].病毒学报,2002,18(2):137-141.
作者姓名:曹胜波  陈焕春  肖少波  赵俊龙  吕建强  钱平
作者单位:华中农业大学畜牧兽医学院病毒室,湖北武汉,430070
基金项目:国家自然科学基金;36170701;
摘    要:参照国外发表的猪Ⅱ型圆环病毒(porcine circovirus type 2,PCV-2)全基因组序列,设计一对PCV-2特异性引物,用该室分离的PCV-2豫A株感染PK-15细胞,从中提取PCV-2复制型基因组DNA,并以之为模板进行PCR扩增.回收PCR产物,构建重组测序质粒T-PCV-2.测序结果表明,猪Ⅱ型圆环病毒豫A株的全基因组为1767bp,与GenBank收录的PCV-2国外分离株核苷酸的同源性可高达97%.序列分析表明,复制型豫A株的基因组包含10个读码框架,其中ORF1、ORF2是其两个最主要的读码框架,分别编码314、234个氨基酸.豫A株和PCV-1间的ORF1、ORF2的氨基酸序列同源性分别为85%、66%,与其它PCV-2毒株间的ORF1氨基酸同源性均在98%以上,而ORF2的氨基酸同源性为92%~97%.

关 键 词:猪Ⅱ型圆环病毒(PCV-2)  基因组  克隆  序列分析
文章编号:1000-8721(2002)02-0137-05

Cloning and Sequencing of Complete Genome of Porcine Circovirus Type 2(PCV-2)YU A Strain
CAO Sheng-bo,CHEN Huan-chun,XIAO Shao-bo,ZHAO Jun-long,LU Jian-qiang,QIAN Ping.Cloning and Sequencing of Complete Genome of Porcine Circovirus Type 2(PCV-2)YU A Strain[J].Chinese Journal of Virology,2002,18(2):137-141.
Authors:CAO Sheng-bo  CHEN Huan-chun  XIAO Shao-bo  ZHAO Jun-long  LU Jian-qiang  QIAN Ping
Abstract:The replicative genomic DNA of porcine circovirus type 2(PCV-2)of YU A strain was extracted from the infected PK-15 cells The complete genome of YU A strain was amplified by utilizing an inverse PCR approach with one pair of special primers. A 1 76kb fragment was obtained and cloned into pMD-18T vector resulting the recombinant plasmid T-PCV-2 that contained the complete genomic DNA of YU A strain The sequence analysis showed that YU A strain genome consisted of 1767nt,which shares 97% sequence identity with other PCV-2 strains in GenBank We also found replicative genome of YU A strain contained ten open reading frames(ORFs) The major ORFs were ORF1 and ORF2,that encoded 314 and 234 amino acids respectively Amino acid sequence homology of ORF1 and ORF2 between YU A strain and PCV-1 were 85% and 66% respectively The ORF1 of YU A strain displayed 98%-99% identity based on amino acids with other PCV-2 strains,but 92%-97% in ORF2
Keywords:porcine  circovirus  type  2(PCV-2)  genome  cloning  sequence  analysis
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