| 摘要: |
| 根据GenBank公布的猪瘟病毒(Classical swine fever virus,CSFV)Shimen株全基因序列,设计并合成2对引物,用RT-PCR方法对采自陕西省部分地区的30份CSF病料中,CSFV流行毒株的E2基因进行了扩增,并将其克隆入pMD18-T载体,转化感受态大肠杆菌DH5α。提取pMD-18T-E2重组质粒,进行BamHⅠ、HindⅢ双酶切鉴定,并对阳性质粒进行测序,将测序结果与HCLV疫苗株和shimen株E2基因及E2蛋白进行序列分析。结果表明,从30份病料中共扩增到10株CSFV流行毒株的E2基因;扩增的10株陕西CSFV流行毒株E2基因同源性为92.3%～99.6%,与Shimen株及HCLV疫苗株的同源性为75.0%～79.4%;10株CSFV流行株E2蛋白的同源性为90.0%～100%,与Shimen株和HCLV疫苗株E2蛋白的同源性分别为78.9%～84.4%和77.8%～83.3%。说明所扩增的10株陕西CSFV流行毒株的E2基因发生了较大变异。 |
| 关键词: 猪瘟 流行毒株 E2基因 序列分析 |
| DOI： |
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| 基金项目:陕西省科技攻关项目（2003KOZ-G11-O3） |
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| Cloning and sequence analysis of E2 gene of ten CSFV strains in Shaanxi |
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| Abstract: |
| Two pairs of special primers were designed and synthesized according to full sequences of CSFV Shimen strain published in GenBank.E2 gene of ten CSFV Shaanxi strains was amplified by reverse transcription polymerase chain reaction(RT-PCR).The PCR product was cloned into pMD-18T vector, then transformed into E.coli DH5α competent cells.The pMD-18T-E2 recombinant plasmid was extracted and the positive plasmid was sequenced after identified by digestion of BamHⅠand Hind Ⅲ restriction enzyme.The results show that nucleotide homogeneity of E2gene of ten CSFV Shaanxi strains is 92.3%-99.6%.Nucleotide homogeneity of E2 gene of ten CSFV Shaanxi strain with E2gene of Shimen and HCLV strain ranged from 75.0%-79.4%.Amino acid homogeneity of E2gene of ten CSFV Shaanxi strains is 90.0%-100%,while 78.9%-84.4% with Shimen strain and 77.8%-83.3% with HCLV strain.The results suggested that mutation occurred in E2 gene of these ten CSFV Shaanxi strains. |
| Key words: classical swine fever,inflowence strain,E2 gene,sequence analysis |
