绵羊IFN-γ基因的克隆与序列分析  

Cloning and Sequence Analysis of Ovine Interferon-γ Gene

作　　者：夏伦斌[1] 连宏军[2] 王新华[2] 闫丽辉[3] 薄新文[2] 钟发刚[2] 荣光[1] 王运宏[1] 郭燕[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832000 [2]新疆农垦科学院兵团绵羊繁育生物技术重点实验室,新疆石河子832000 [3]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001

出　　处：《动物医学进展》2006年第10期69-72,共4页Progress In Veterinary Medicine

基　　金：新疆生产建设兵团绵羊繁育生物技术重点实验室开放课题(05KLS10)

摘　　要：将无菌采集的绵羊(湖羊)脾脏淋巴细胞进行体外培养,利用刀豆蛋白(ConA)刺激24 h后提取总RNA,采用RT-PCR扩增绵羊IFNγ-基因,并将其克隆到pMD18-T载体中,经PCR和双酶切鉴定后进行测序。测序结果表明,扩增的cDNA全长554 bp,包含501 bp开放阅读框,编码166个氨基酸,分子质量约为18.4 ku。克隆的绵羊IFNγ-基因与GenBank上已公布的人、牛、山羊、马鹿、马、骆驼、猪、狗、猫和鸡的IFNγ-核苷酸序列进行比较,其同源性分别为75.2%,96.6%,99.0%,95.8%,83.0%,88.8%,86.6%,82.4%,53.9%和32.9%,与其他品种绵羊的核苷酸同源性为100%;氨基酸序列同源性分别为61.7%,94.6%,98.2%,92.2%,77.8%,86.8%,77.8%,76.0%,39.5%和6.7%,这为进一步研究绵羊IFNγ-基因的表达、生物学活性和应用奠定了一定基础。Spleen lymphocytes isolated from sheep were cultivated in RPMI-1640 culture medium, then concavadin A was used to stimulate the spleen lymphocytes for 24h,and then the total RNA isolated from the lymphocyte, the ovine IFN-γ gene was amplified with RT-PCR. The gene segment of IFN-γ was cloned into the vector pMD18-T,and then sequenced. The result suggested that the full length sequence of amplifying ovine IFN-γ gene cDNA consisted of 554bp,which contained a complete open reading frame of 501 bp, it encoded 166 amino acids. Compared with the published IFN-γ gene sequence, the homology of nucleotide sequence was 75.2% with human,96.6% with sheep,99.0% with goat,95.8% with red deer,83.0% with horse,88.8% with camel,86.6% with pig,82.4% with dog,53.9% with cat and 32.9% with chicken;The homology of IFN-γ was 100% at nucleotide level between Hu sheep and other breed sheep. This study established the way for future study on biological function,activity and application of ovine IFN-γ.

关 键 词：绵羊 IFN-Γ基因 克隆 序列分析 

分 类 号：Q78[生物学—分子生物学] S858.26[农业科学—临床兽医学]