乌鬃鹅α干扰素基因的克隆及生物信息学分析

CLONING AND BIOINFORMATICS ANALYSIS OF IFN-α GENE OF WUZONG GOOSE

采用RT-PCR方法从乌鬃鹅肝脏中扩增α干扰素(Interferon,IFN-α)基因片段,将扩增片段克隆到pMD18-T载体上,经PCR和双酶切鉴定为阳性后进行序列测定,并用生物信息学软件Lasergene v7.1 DNAStar和在线分析方法对IFN-α基因的核苷酸序列及其氨基酸序列进行了生物信息学分析,同时与GenBank中登录的鸡、鸭和鹅IFN-α基因核苷酸序列进行同源性比较分析。结果表明,所扩增的IFN-α基因编码完整的开放阅读框,基因长度为576 bp,与鸡、鸭与鹅IFN-α基因核苷酸序列同源性在72.0%～99.7%,遗传进化树显示鸡、鸭、鹅IFN-α因在遗传进化上各处一支。乌鬃鹅IFN-α基因的克隆及生物信息学分析为进一步研究鹅干扰素抗病毒的作用机理奠定基础。

A IFN-a gene was amplified from the liver tissue of Wuzong goose using RT-PCR and was cloned into the vector pMD18-T -the positive clones were confirmed by PCR and restriction enzyme digestion. The nucleotide sequences of the amplified IFN-a and the amino acid sequences deduced from the gene were the analyzed by the bioinformatice software Lasergene v7.1 DNAStar and on-line tools. The similarity and evolution of IFN-a gene were analyzed among Wuzong goose and otherl5 species of poultry （chicken, duck and goose）. These results revealed that the cloned-successfully IFN-a gene shared 72.0%-99.7% similarity with that of the 15 species of poultry, and they had it＇s own independent branch in the phylogenetic tree. The study laid the foundations for the further study of mechanism of goose interferon antivirus.