坝上长尾鸡MITF基因核心启动子鉴定与分析

doi:10.11843/j.issn.0366-6964.2018.06.003

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (6): 1116-1123.doi: 10.11843/j.issn.0366-6964.2018.06.003

坝上长尾鸡MITF基因核心启动子鉴定与分析

刘小辉^1,2, 彭永东², 周荣艳¹, 张传生², 李祥龙^1,2*

1. 河北农业大学动物科技学院, 保定 071001;
2. 河北科技师范学院动物科技学院, 秦皇岛 066004

收稿日期:2017-11-28 出版日期:2018-06-23 发布日期:2018-06-22
通讯作者: 李祥龙,教授,E-mail:lixianglongcn@yahoo.com
作者简介:刘小辉(1988-),女,河北邯郸人,博士生,主要从事动物遗传育种研究,E-mail:liuxiaohui1000@163.com
基金资助:
河北省蛋鸡体系产业技术创新团队（HBCT2013090206）；河北省科技计划项目（15226302D）

Identification and Analysis of the Core Promoter of MITF Gene in Bashang Long-tail Chicken

LIU Xiao-hui^1,2, PENG Yong-dong², ZHOU Rong-yan¹, ZHANG Chuan-sheng², LI Xiang-long^1,2*

1. College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, China;
2. College of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066004, China

Received:2017-11-28 Online:2018-06-23 Published:2018-06-22

摘要/Abstract

摘要：

旨在探明坝上长尾鸡小眼畸形相关转录因子（microphthalmia-associated transcription factor，MITF）基因启动子活性区与结构，为研究MITF基因在毛囊组织中的表达调控提供依据。本研究采用双荧光素酶表达载体的方法，构建了6个含有不同长度坝上长尾鸡MITF基因启动子片段的pGL3-Basic表达载体及4个核心启动子区突变载体，转染DF1细胞48 h后检测双荧光素酶活性。结果，确定了鸡MITF基因启动子的核心区域为-660~+200 bp。其中突变位点-579 bp、-505 bp、-274 bp、-220 bp、-203~-202 bp、-98 bp、-46 bp、-14 bp、+1 bp、+28 bp对MITF基因启动子活性有较大影响，在突变区域预测到HOX家族（HOXA3、HOXD8、HOXD9、HOXD10、HOX11）、NF-1、DBP、TFⅡD和TFⅡB与MITF序列的结合性发生了变化。

关键词: 坝上长尾鸡, MITF基因, 启动子, 转录因子

Abstract:

The research was designed to study the activity and the structure of MITF promoter and to provide clues for studying this gene's regulation mechanism in hair follicle. Dual-luciferase expression vectors were constructed and transfected to DF1 cells with lip2000. The dual-luciferase detection kit was used to measure the relative luciferase activity. The 6 expression vectors with different promoter regions of MITF gene and 4 mutant vectors of the core promoter region were constructed in Bashang Long-tail chicken. The core promoter region from -660 bp to +200 bp was identified in chicken MITF gene. The function of -579 bp, -505 bp, -274 bp, -220 bp, -203--202 bp, -98 bp, -46 bp, -14 bp, +1 bp and +28 bp sites in regulating promoter activity was critical. The similarities of transcription factor binding sites including HOX family (HOXA3, HOXD8, HOXD9, HOXD10 and HOX11), NF-1, DBP, TFⅡD and TFⅡB to MITF were changed predicted by software.

Key words: Bashang Long-tail chicken, MITF gene, promoter, transcription factor

中图分类号:

S831.2

刘小辉, 彭永东, 周荣艳, 张传生, 李祥龙. 坝上长尾鸡MITF基因核心启动子鉴定与分析[J]. 畜牧兽医学报, 2018, 49(6): 1116-1123.

LIU Xiao-hui, PENG Yong-dong, ZHOU Rong-yan, ZHANG Chuan-sheng, LI Xiang-long. Identification and Analysis of the Core Promoter of MITF Gene in Bashang Long-tail Chicken[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1116-1123.

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坝上长尾鸡MITF基因核心启动子鉴定与分析

Identification and Analysis of the Core Promoter of MITF Gene in Bashang Long-tail Chicken

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