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河北省猪圆环病毒2型ORF2基因的克隆与序列分析
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李鹏1， 郑其升1，李斐1，任雪枫2，仇妍虹1，牛明福1，周斌1，
曹瑞兵1，陈溥言1
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(1.南京农业大学 农业部动物疫病诊断与免疫重点开放实验室，江苏 南京210095；
2.江苏省畜牧兽医站,江苏 南京210036)
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摘要：
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根据GenBank 中猪圆环病毒2 型( PCV2)ORF2 基因的核苷酸序列,设计了1对特异性引物。采用此引物从PCV2疑似病料的3个样品中扩增出了ORF2片段。将扩增片段克隆入pMD18-T载体中,筛选获得了含有相应片段的阳性重组质粒pMDHB-ORF2。对质粒中的插入片段测序后，应用DNAStar序列分析软件,对所测PCV2 序列与GenBank 中的国内外PCV毒株进行了同源性比较。结果，3个样品间ORF2基因的核苷酸同源性为100%；它们与浙江分离株之间核苷酸序列的同源性极高,达99.5%。 |
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关键词：
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猪圆环病毒2型；ORF2基因；克隆；序列分析 |
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中图分类号：S 852.659.2：Q 503 文献标识码：
A 文章编号：1673-4696(2006)04-0262-04 |
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Cloning and sequence analysis of ORF2 gene of porcine
circovirus type 2 from Hebei Province |
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LI Peng1, ZHENG Qi-sheng1, LI Fei1, REN
Xue-feng2, QIU Yan-hong1, NIU Ming-fu1，
ZHOU Bin1, CAO Rui-bing1, CHEN Pu-yan1 |
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(1. Key Laboratory of Animal Disease Diagnosis and Immunology, Nanjing Agricultural University, Nanjing
210095,China; 2.Animal Husbandry and Veterinary Station of Jiangsu Province, Nanjing 210036, China) |
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Abstract: Using a pair of specific primers designed according to the relevant sequences from GenBank, ORF2 genes of three porcine circovirus type 2 specimens from suspected pigs in Hebei Province were amplified by PCR, respectively. The amplified fragments were cloned into the pMD18 T vectors and the positive recombinant plasmids were designated pMDHB ORF2. After sequencing, the ORF2 genes were compared with PCV2 genome sequences downloaded from GenBank by DNAStar software and then the phylogenetic tree was constructed. The results revealed that ORF2 gene sequences of the three specimens were identical, and the similarity between the specimens and the isolated strain from Zhejiang Province was 995%. |
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Key words: porcine circovirus type 2; ORF2 gene; cloning; sequence analysis |
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