[1]陈哲,雷明明,于建宁,等.猪RELMβ基因启动子区克隆及序列分析[J].江苏农业学报,2015,(05):1060-1064.[doi:doi:10.3969/j.issn.1000-4440.2015.05.018]
　CHEN Zhe,LEI Ming-ming,YU Jian-ning,et al.Cloning and sequence analysis of promoter region of porcine RELMβ gene[J].,2015,(05):1060-1064.[doi:doi:10.3969/j.issn.1000-4440.2015.05.018]
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猪RELMβ基因启动子区克隆及序列分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年05期
页码:
1060-1064
栏目:
畜牧兽医·水产养殖
出版日期:
2015-10-31

文章信息/Info

Title:
Cloning and sequence analysis of promoter region of porcine RELMβ gene
作者:
陈哲; 雷明明; 于建宁; 王公金; 施振旦
（江苏省农业科学院畜牧研究所，动物品种改良与繁育重点实验室,江苏南京210014）
Author(s):
CHEN Zhe; LEI Ming-ming; YU Jian-ning; WANG Gong-jin; SHI Zhen-dan
(Key Laboratory of Animal Breeding and Reproduction, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
猪; 抵抗素样β基因; 启动子
Keywords:
swine; RELMβ; promoter
分类号:
S828
DOI:
doi:10.3969/j.issn.1000-4440.2015.05.018
文献标志码:
A
摘要:
本研究旨在分析猪RELMβ基因启动子结构，初步探索RELMβ基因表达调控机制。通过PCR方法扩增RELMβ基因的系列启动子缺失片段并分别克隆到荧光素酶报告基因表达载体pGL3-Enhancer中，经酶切、测序和生物信息学分析，构建包含RELMβ启动子系列截短的荧光素酶报告基因重组质粒，脂质体转染至HT29和293T细胞，应用双荧光素酶活性检测系统检测启动子活性。试验获得了猪RELMβ基因约1 kb的启动子序列，序列比对发现猪和人物种间相似性仅34.9%，猪和小鼠的同源性是82.4%。生物信息学分析预测猪RELMβ基因转录起始位点在-556 bp处，猪和人RELMβ基因启动子存在系列保守的转录因子结合位点，包括Cdx2、SRY、NFKB、NKX-2、c-Myb、GATA-1、GATA-3、C/EBP、MZF1等。细胞检测结果显示，pGL-RELMβ（-574～+215）活性最强，推测在-574～-182 bp位点之间存在RELMβ基因启动子的关键顺式调控元件，这一区域发现SRY、Cdx2、GATA-1和MZF1等关键转录因子结合位点。
Abstract:
The aim of this study was to identify the promoter structure of porcine RELMβ gene,and investigate its transcriptional regulation mechanism. The deletion fragments of porcine RELMβ gene promoter region were amplified by PCR method, cloned into pGL3-Enhancer plasmid by digestion, sequencing and bioinformatics analysis, and transfected into HT29 and 293T cells. The promoter activities were determined by dual-luciferase assay system. An approximate 1 kb promoter sequence was acquired and the homologies were 34.9% and 82.4% to RELMβ gene of human and mouse, respectively. Bioinformatics analysis found a potential transcription start site located at -556 bp, and the conservative transcription factor binding sites in pig and human RELMβ gene included Cdx2, SRY, NFKB, NKX-2,c-Myb, GATA-1, GATA-3, C/EBP and MZF1. pGL-RELMβ（-574-+215） exhibited the maximal promoter activities, suggesting the region of -574--182 bp contained the key cis-regulatory elements. Further bioinformatics analysis found some key transcription factor binding sites, including SRY, Cdx2, GATA-1 and MZF1.

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备注/Memo

备注/Memo:
收稿日期：2015-02-12 基金项目：江苏省农业科技自主创新基金项目[CX(12)5010]；江苏省自然科学基金项目（BK20140753）；转基因生物新品种培育重大专项（2014ZX08006-004） 作者简介：陈哲（1982-），男，山东泰安人，博士，助理研究员，主要从事动物遗传育种研究。（Tel）025-84390341；（E-mail）chenzzju@163.com 通讯作者：施振旦，（Tel）025-84390956；（E-mail）zdshi@mail.jaas.ac.cn
更新日期/Last Update: 2015-10-31