| 摘要: |
| 根据GenBank中已发表的鸡传染性支气管炎病毒(IBV)N基因序列,设计并合成1对特异性引物,用RT-PCR方法扩增IBVHN99株的N基因,并将其克隆入pMD18-T载体,转化感受态大肠杆菌Top10,提取pMD18-T-N重组质粒,进行酶切和PCR鉴定,并对阳性质粒进行测序,将测序结果与H52,Ark99,BJ等参考毒株进行序列分析。结果表明,N基因全长为1230bp,编码1条409个氨基酸组成的多肽;其核苷酸与H52,Ark99和BJ等毒株的同源性为88.4%～90.7%,氨基酸同源性为91.7%～94.4%;HN99株与H52,Ark99和BJ等其他各毒株的亲缘关系均较远,是1株新的IBV变异株。 |
| 关键词: 鸡传染性支气管炎病毒 N基因 基因克隆 序列分析 |
| DOI： |
| 分类号: |
| 基金项目:河南省科技攻关项目(0424050015) |
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| Cloning and sequence analysis of N gene of infectious bronchitis virus HN99 strain |
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| Abstract: |
| A pair of special primers were designed and synthesized according to avian infectious bronchitis virus N gene sequences published in GenBank.N gene of IBV HN99 strain was amplified by reverse transcription-polymerase chain reaction (RT-PCR).The PCR product was cloned into pMD-18T vector,then transformed into E.coli Top10 competent cells.The pMD-18T-N recombinant plasmid was extracted and the positive plasmid was sequenced after identification by digestion of restriction enzyme and PCR.The results show that cDNA of N gene of HN99 was composed of 1 230 nucleotides encoding a polypeptide of 409 amino acids.Nucleotide homogeneity of N gene of HN99 with N gene of other IBV strains H52,Ark99,BJ ranged from 88.4-90.7,while 91.7-94.4 identity was found on amino acid level.Evolution analysis indicated that HN99 was far related to H52,Ark99,BJ and other IBV strains.These results suggested that IBV HN99 was a new variant of IBV. |
| Key words: avian infectious bronchitis virus N gene genetic cloning sequence analysis |
