鹌鹑和三黄鸡IFN-α cDNA的克隆及序列分析  

Molecular Cloning and Sequencing Analysis of the Alpha Interferon from Chicken and Quail

作　　者：汪镇南[1] 苏丁丁[2] 刘为[3] 王亚[1] 肖利军[1] 李润成[1] 白霞[1] 余兴龙[1] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128 [2]湖南农业大学动物科学技术学院,湖南长沙410128 [3]湖南师范大学生命科学院,湖南长沙410081

出　　处：《激光生物学报》2008年第3期367-370,共4页Acta Laser Biology Sinica

基　　金：湖南省科技厅重大项目(2006NK2003)

摘　　要：根据GenBank已登录的鸡和鹌鹑IFN-α基因序列,用Oligo6.0设计一对能同时扩增鸡和鹌鹑INF-α基因的共用引物,利用RT-PCR方法从经ConA诱导培养的三黄鸡、鹌鹑外周血淋巴细胞中分别扩增到IFN-α ORF全长cDNA,并克隆到pET-28a（＋）载体上。将测序结果分别与GenBank中已收录的家禽类IFN-α核苷酸序列进行同源性比较及遗传进化分析。结果发现鸡、鹌鹑IFN-α cDNA遗传进化稳定,与GenBank中收录的鸡、鹌鹑IFN-α核苷酸序列同源性为98.6%-99.8%、99.8%。鸡与其它家禽类的IFN-α核苷酸序列比较分析发现,与鹌鹑亲缘性较高,同源性达86.9%-87.1%,但与家水禽（鸭、鹅）差距较大,同源性最高也只有67.6%;鹌鹑IFN-α核苷酸序列与家水禽同源性在也只在62.7%-62.9%之间;基因遗传进化树分析可见家禽类IFN-α被分成两个大分枝,分别是由鸡和鹌鹑两个小分支组成的一个大分支与家水禽独立分支组成。One pair of primers were designed according to chicken and quail IFN-α gene sequences published in Gen- Bank. And were used to amplify chicken and quail IFN-α eDNA by RT-PCR from total RNA of chicken and quail lymphocytes induced by ConA. RT-PCR product was cloned into pET-28a（ ＋ ） vector. Sequence was sequenced, subtyped and put up homologous analysis. The results suggested that the chicken and quail IFN-α gene were very conservative, compared with the nucleotide sequence registered in GenBank, the homology of chicken and quail IFN-α were 98.6 % - 99.8 % and 99.8 %. Chicken Compared with other pouhry, the IFN-α nucleotide sequence analysis showed that the relationship of quail had high homology of 86.9 % - 87.1%, but it had big disparity from aquatic bird （ ducks and geese） , and the highest homology was only 67.6 percent; the nucleotide sequence of IFN-α of quail homology compared with the domestic birds was only 62.7 % -62.9 %. The analysis of genetic tree showed that the IFN-α poultry was divided into two large branches, the one were chickens and quails, the other were the aquatic bird. .

关 键 词：鸡 鹌鹑 IFN-Α 基因克隆 序列分析 

分 类 号：Q523[生物学—生物化学]