牛FATP1基因5&apos;侧翼启动子克隆及序列分析

生物技术通报 ›› 2013, Vol. 0 ›› Issue (5): 86-92.

牛FATP1基因5'侧翼启动子克隆及序列分析

龚婷, 许厚强, 陈伟, 赵佳福, 骆衡, 陈祥, 张勇

(1. 贵州大学高原山地动物遗传育种与繁殖省部共建教育部重点实验室，贵阳 550025；2. 贵州大学贵州省动物遗传育种与繁殖重点实验室，贵阳 550025)

收稿日期:2012-12-27 修回日期:2013-05-24 出版日期:2013-05-24 发布日期:2013-05-24
作者简介:龚婷，女，硕士研究生，研究方向：动物遗传育种与繁殖；E-mail：750980455@qq.com
基金资助:
国家转基因生物新品种培育科技重大专项(2011ZX08009-004)，贵州省科技计划课题[黔科NY字(2012)3008号]

Cloning and Sequence Analysis of 5'-Flanking Region of Bovine FATP1 Gene

Gong Ting, Xu Houqiang, Chen Wei, Zhao Jiafu, Luo Heng, Chen Xiang, Zhang Yong

(1. Key Laboratory of Animal Genetics，Breeding and Reproduction in the Plateau Mountainous Region，Guizhou University，Guiyang 550025；2. Ministry of Education Key Laboratory of Animal Genetics，Breeding and Reproduction，Guizhou University，Guiyang 550025)

Received:2012-12-27 Revised:2013-05-24 Published:2013-05-24 Online:2013-05-24
About author:许厚强，男，博士，教授，博士生导师，研究方向：分子遗传与动物育种；E-mail：houqiang0524@yahoo.com

摘要/Abstract

摘要： 旨在克隆牛FATP1基因5'侧翼启动子，研究其特异性转录调控元件的调控机制。利用PCR方法扩增牛FATP1基因5'侧翼区，并通过产物纯化、连接、转化及测序比对，获得2 164 bp (-1 969- +194 bp)的5'侧翼启动子序列。利用生物信息软件对获得的2 164 bp 5'侧翼启动子克隆载体进行分析预测，发现其有4处转录起始点和40个潜在转录因子结合位点；该序列与人、小鼠、大鼠和狗的同源性分别为74.1%、71.1%、72.0%、62.8%，且不同物种FATP1基因5'侧翼区的同源序列主要集中在转录起始点上游-578- -93 bp区域，保守性较强的区域在转录起始点上游-263- -174 bp，推测转录起始点上游-263- -174 bp可能是其核心启动子区域，从而成功克隆了FATP1 5'侧翼启动子序列2 164 bp，初步预测了该基因的核心启动子区域。

关键词: 牛, 克隆, FATP1基因, 启动子, 转录因子

Abstract: The experiment was conducted to clone 5'-flanking region of bovine FATP1 gene to study regulatory mechanism of the specific transcriptional regulation region. The 5'-flanking region of bovine FATP1 gene was amplified with PCR, and then the 2 164 bp (-1969- +194 bp) 5'-flanking promoter sequence was obtained by product purification, ligation, transformation, and sequence comparison. The obtained cloning vector of 5'-flanking region of bovine FATP1 gene was analyzed by the promoter software in this study. The results indicated that there were 4 transcription start sites and 40 potential transcription factor binding sites. Comparing bovine with human, mouse, rat and dog, the homology of 5'-flanking promoter sequence were 74.1%, 71.1%, 72.0%, and 62.8%, respectively. The conservative region was mainly concentrated in -578- -93 bp of the upstream of transcription start sites, and the highly conserved region was in -265- -162 bp of the upstream of transcription start sites, which may be conclude that the -265- -162 bp of the upstream of transcription start sites was the core promoter region of the gene. The 2 164 bp 5'-flanking region of the bovine FATP1 gene was cloned successfully and the core promoter region was preliminary predicted in the gene.

Key words: Bovine, Cloning FATP1 gene, Promoter, Transcription factor

龚婷, 许厚强, 陈伟, 赵佳福, 骆衡, 陈祥, 张勇. 牛FATP1基因5'侧翼启动子克隆及序列分析[J]. 生物技术通报, 2013, 0(5): 86-92.

Gong Ting, Xu Houqiang, Chen Wei, Zhao Jiafu, Luo Heng, Chen Xiang, Zhang Yong. Cloning and Sequence Analysis of 5'-Flanking Region of Bovine FATP1 Gene[J]. Biotechnology Bulletin, 2013, 0(5): 86-92.

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(责任编辑李楠)

牛FATP1基因5'侧翼启动子克隆及序列分析

Cloning and Sequence Analysis of 5'-Flanking Region of Bovine FATP1 Gene

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