摘要： 基于电子延伸序列,克隆并分析了绵羊Gastorkine-1基因。提取皱胃黏膜组织总RNA,利用设计的引物进行RT-PCR,PCR产物与pMD19-T载体连接后转化E.coliJM109感受态细胞,检测阳性克隆并测序。克隆的绵羊Gas-torkine-1基因长619 bp,编码185个氨基酸,与人、小鼠、猪、牛的同源性分别为82.0%,48.8%,85.4%,96.9%,推测的氨基酸序列信号肽为1~20 aa,BRICHOS结构域为54~150 aa,结构特征与人、小鼠、猪、牛的Gastorkine-1相一致。
关键词:
绵羊,
胃黏膜保护因子,
克隆,
序列分析
Abstract: In order to clone and analysis the ovine Gastorkine -1, the known sequence of bovine Gastorkine -1(NM001001148)was selected to search the GenBank ovine EST database, from which two ESTs(EE780384, EE77958)were obtained. Based on the sequence spliceosome, a pair of primers in 5 c -UTR and 3c -UTR was designed and used toamplify the total RNA extracted from mucosal tissue of ovine abomasum by RT -PCR,then the PCR products were clonedand sequenced.The results showed that ovine Gastorkine -1 sequence shared 8210%, 4818%, 8514% and 9619% sim- ilarity with the corresonding genes of human,mouse, pig and cattle, respectively.The signal peptide of predicted amino acidsequence contained 1- 20 aa and the structural domain BRICHOS had 54- 150 aa, presenting the same structural featureto those of human,mouse, pig and cattle.
Key words:
Sheep,
Gastorkine -1,
Molecular cloning,
Sequence analysis
中图分类号:
乔新安, 杨国宇, 王艳玲, 王月影, 韩立强, 朱彦彩, 都军霞, 范沛. 绵羊Gastorkine-1基因的克隆与序列分析[J]. 华北农学报, 2008, 23(1): 38-40. doi: 10.7668/hbnxb.2008.01.009.
QIAO Xin-an, YANG Guo-yu, WANG Yan-ling, WANG Yue-ying, HAN Li-qiang, ZHU Yan-cai, DU Jun-xia, FAN Pei. Molecular Cloning and Sequence Analysis of Ovine Gastorkine-1[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(1): 38-40. doi: 10.7668/hbnxb.2008.01.009.