犬白细胞介素18基因的克隆与序列分析

为研究犬白细胞介素18(IL-18)的生物学功能,从犬外周血中分离白细胞,经Con A刺激后,提取总RNA,通过反转录-聚合酶链反应(RT-PCR)扩增犬IL-18基因,连接pMD19-T simple vector,转化DH5α感受态细胞,经双酶切鉴定,获得阳性重组质粒后进行序列分析。结果表明,获得的犬IL-18基因全长为582bp,编码氨基酸194个。将犬与GenBank中牛、猫、羊、猪、鼠、兔、狐、貉IL-18基因进行同源性比较,发现犬IL-18与红狐和貉IL-18的同源性最高,氨基酸序列同源性分别达96.4%、91.2%,与其他物种则存在较大种属差异。

Cloning and Sequence Analysis of Canine Interleukin-18 Gene

To laid the foundation for a further study and investigation of the function of canine IL-18,the IL-18 gene was amplified by RT-PCR from the total RNA of canine peripheral blood leucocyte induced with Con A.The RT-PCR product was cloned into pMD19-T simple vector.The nucleotide sequence had been determined after being identified by double restriction enzyme digestion.The results indicated that clone canine IL-18 gene was 582 bp in length encoding a polypeptide of 193 amino acid.Compared with the gene of Vulpes vulpes,Nyctereutes procyonoides,Sus scrofa,Ovis aries,Bos taurus,Oryctolagus cuniculus,Rattus norvegicus,Felis catus on GenBank,respectively,the aminoacid homology of canine IL-18 showed 96.4% and 91.2% with that of Vulpes vulpes IL-18 and Nyctereutes procyonoides IL-18.There is significant difference between IL-18 gene sequences of dog and other species.