绵羊Granulysin基因的克隆与序列分析  

Analysis on Molecular Cloning and Sequence of Ovine Granulysin

作　　者：乔新安[1] 杨国宇[1] 朱彦彩[2] 王月影[1] 韩立强[1] 王艳玲[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002 [2]河南科技学院动物科学学院,新乡453003

出　　处：《华中农业大学学报》2008年第3期394-397,共4页Journal of Huazhong Agricultural University

基　　金：河南省重点科技攻关项目(0523010500)资助

摘　　要：基于电子延伸序列,设计1对克隆引物,从绵羊回肠黏膜组织中提取总RNA,进行RT-PCR,将PCR产物与pMD19-T载体连接后转化E.coliJM109感受态细胞、检测阳性克隆、测序并进行序列分析。克隆的绵羊Granulysin基因与人、牛该基因的同源性分别为56.2%和87.2%,推导的氨基酸序列信号肽为1～22氨基酸,SapB结构域为64～138氨基酸,结构特征与人、牛的一致。结果提示,绵羊Granulysin基因克隆成功。A pair of cloning primers were designed based on the in silico sequence information. Total RNA was extracted from mucosal tissue of ovine ileum and mRNA sequence was amplified by RT-PCR. The PCR products were ligated into the pMD19-T vector, and then transformed into competent cells of E. coli JM109. The positive clone was identified and the sequence was sequenced and analysed. The results showed that ovine Granulysin shared 56. 2% and 87. 2% homology with that of human and cattle. The signal peptide of deduced protein sequence is 1-22 amino acid and the structural domain SapB is 64-138 amino acid, and the structural feature is consistent with human and cattle. The ovine Granulysin gene was successfully cloned.

关 键 词：绵羊 颗粒溶解素 克隆 序列分析 

分 类 号：S811.3[农业科学—畜牧学] Q789[农业科学—畜牧兽医]