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猪圆环病毒2型山东株全基因组的克隆与序列分析
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柳美玲1，秦晓冰1，王爱华2 ，牛晋国3，单虎1*
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（1.青岛农业大学 动物科技学院，山东 青岛266109；2.云南农业大学 动物科技学院，云南 昆明650201；
3.山西省农业科学院 畜牧兽医研究所，山西 太原030032）
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摘要：
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参照国外发表的猪Ⅱ型圆环病毒（PCV2）全基因组序列,设计合成了1对特异性引物，从山东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，进行PCR扩增。回收PCR产物，将其插入pMD18-T载体，构建了重组质粒pMD18-T-PCV2，并对筛选出的阳性质粒进行测序。结果表明，克隆得到的PCV2山东株的全基因组长为1767bp。应用DNAStar序列分析软件，对所测PCV2序列与GenBank中登录的国内外PCV毒株进行同源性比较。结果显示，PCV2山东株与国内外毒株的核苷酸同源性高达99.6%，推导的氨基酸同源性达95.4%。进化树分析结果显示,PCV2山东株与GZ（EF515839）株同源性最高，表明，PCV2各分离毒株在进化方面存在地域上的相关性。 |
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关键词：
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猪Ⅱ型圆环病毒；基因组；克隆;序列分析 |
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中图分类号：S 852.659.2 文献标识码：
A 文章编号：1673-4696(2008)01-0010-05 |
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Cloning and sequence analysis of the complete genome of
porcine circovirus type 2 Shandong strain |
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LIU Mei-ling1,QIN Xiao-bing1,WANG
Ai-hua2,NIU Jin-guo3，SHAN Hu1 |
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(1.College of Animal Science and Technology,Qingdao Agricultural University,Qingdao 266109,China;
2.College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;
3.Husbandry and Veterinary Institute,Shanxi Academy of Agricultural Sciences,Taiyuan 030032,China) |
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Abstract: According to the published genome sequences of PCV2, one pair of specific primers were designed to amplify the full length of viral genome from tissues of clinical pigs suspected to be post weaning multisystemic wasting syndrome(PMWS) in Shandong Province.The amplified fragment was cloned into pMD18-T vector to obtain the recombinant plasmid pMD18-T-PCV2. Sequence analysis showed that the genome of PCV2 Shandong(SD) strain consisted of 1767bp,which shared 99.6% sequence identity and 95.4% amino acid sequence identity with other PCV-2 strains in GenBank. Phylogenetic analysis showed that the SD strain shared the highest sequence identity with GZ strain（EF515839） and there was a geographical correlation among the PCV2 isolates. |
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Key words: porcine circovirus type 2( PCV-2);genome;cloning;sequence analysis |
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