关岭牛解偶联蛋白3基因5’侧翼区克隆和生物信息学分析

中国畜牧兽医 ›› 2014, Vol. 41 ›› Issue (9): 6-10.

关岭牛解偶联蛋白3基因5’侧翼区克隆和生物信息学分析

陈伟^1,2,3, 许厚强^1,2, 刘忠伟³, 陈祥^1,2, 张雯^1,2, 桓聪聪^1,2,3

1. 贵州大学动物科学学院, 贵州贵阳 550025;
2. 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, 贵州贵阳 550025;
3. 贵州大学生命科学学院, 贵州贵阳 550025

修回日期:2014-06-11 出版日期:2014-09-20 发布日期:2014-09-24
通讯作者: 许厚强 E-mail:houqiang0524@yahoo.com
作者简介:陈伟（1980- ），女，内蒙古人，博士生，研究方向：遗传学。
基金资助:
国家转基因生物新品种培育科技重大专项（2011ZX08009-004）；贵州省科技厅农业攻关计划（黔科NY字[2012]3008号）；贵州省科学技术基金项目（黔科合J字[2012]2145号）。

Cloning and Sequence Analysis of UCP3 Gene 5’-Flanking Region of Guanling Cattle

CHEN Wei^1,2,3, XU Hou-qiang^1,2, LIU Zhong-wei³, CHEN Xiang^1,2, ZHANG Wen^1,2, HUAN Cong-cong^1,2,3

1. College of Animal Science, Guizhou University, Guiyang 550025, China;
2. Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Guizhou University, Guiyang 550025, China;
3. College of Life Science, Guizhou University, Guiyang 550025, China

Revised:2014-06-11 Online:2014-09-20 Published:2014-09-24

摘要/Abstract

摘要： 本试验对贵州关岭牛解偶联蛋白3（uncoupling protein，UCP3）基因5'侧翼区进行克隆，并利用生物信息学软件对其1080 bp的克隆序列进行分析，以研究UCP3基因5'侧翼区特异性转录调控元件的调控机制。软件分析发现关岭牛UCP3基因5'侧翼区含有6个可能的转录起始点和33个潜在转录因子结合位点，与东北虎、家猫、印度水牛、藏羚羊、山羊、绵羊UCP3基因5'侧翼区（-196～+100 bp）序列相似性分别为82%、82%、98%、95%、96%和98%；该区域保守性较强，推测转录起始点上游-200～+1 bp可能是其核心启动子区域，该区域在调控基因转录和翻译方面具有共同的作用。试验成功克隆了UCP3基因5'侧翼区序列1080 bp，初步预测了该基因的核心启动子区。

关键词: 关岭牛; 解偶联蛋白3基因; 启动子; 转录因子

Abstract: The experiment was conducted to clone 5'-flanking region of bovine UCP3 gene to study regulatory mechanism of the specific transcriptional regulation region. The 5'-flanking region of bovine UCP3 gene was amplified with PCR, and then the 1080 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 UCP3 gene was analyzed by the promoter software in this study. The results indicated that there were 6 transcription start sites and 33 potential transcription factor binding promoter region of the gene, the highly conserved region was in -196～+100 bp of the upstream of transcription start sites, which might be conclude that the -200～+1 bp of the upstream of transcription start sites was the core. Comparing bovine with Panthera tigris, Felis catus, Bubalus bubalis, Pantholops hodgsonii, Capra hircus, Ovis aries,the homology of 5'-flanking promoter sequence were 82%, 82%, 98%, 95%, 96% and 98%, respectively. The 1080 bp 5'-flanking region of the bovine UCP3 gene was cloned successfully and the core promoter region was preliminary predicted in the gene.

Key words: Guanling cattle; UCP3 gene; promoter; transcription factors

中图分类号:

Q785

陈伟, 许厚强, 刘忠伟, 陈祥, 张雯, 桓聪聪. 关岭牛解偶联蛋白3基因5’侧翼区克隆和生物信息学分析[J]. 中国畜牧兽医, 2014, 41(9): 6-10.

CHEN Wei, XU Hou-qiang, LIU Zhong-wei, CHEN Xiang, ZHANG Wen, HUAN Cong-cong. Cloning and Sequence Analysis of UCP3 Gene 5’-Flanking Region of Guanling Cattle[J]. , 2014, 41(9): 6-10.

参考文献

1 文生萍,张丽春,曹振辉,等. 解偶联蛋白及其基因多态性与能量代谢[J].中国畜牧兽医, 2008, 35(4): 42～45.

2 刘敏, 许厚强,陈伟,等.关岭黄牛MSTN启动子真核报告载体的构建与启动活性研究[J].广东农业科学,2013, 40(17):133～136.

3 闫晓红,王宁. 转录因子与microRNA在基因表达调控中的功能联系及差异[J]. 中国生物化学与分子生物学报,2010,10:892～897.

4 张润锋,陈宏,支立峰,等. 南阳牛IGF-ⅠR、CAPNⅡ和UCP3基因多态性与生长性状的相关分析[J].农业生物技术学报,2010,18 (5):893～898.

5 李海莲. UCP3基因的研究进展[J].上海畜牧兽医通讯,2011,3:15～16.

6 陈翠,张立敏,陈晓杰,等. UCP3基因遗传多态性与中国西门塔尔牛生长及育肥性状的遗传效应分析[J].中国农学通报,2012, 28(5): 13～18.

7 罗启华,王安娜. 贵州关岭黄牛发展现状与思考[J].草食家畜,2011(4):15～16.

8 韩凤桐. 牛SRY基因核心启动子定位及其基因功能研究[D].哈尔滨: 东北农业大学, 2008.

9 Boss O, Samec S, Dulloo A, et al. Tissue-dependent upregulation of rat uncoupling protein-2 expression in response to fasting or cold [J]. FEBS Lett, 1997, 412(1): 111～114.

10 Chakravarthy M V, Lodhi I J, Yin L, et al. Identification of a physiologically relevant endogenous ligand for PPAR alpha in liver[J]. Cell, 2009, 138(3):476～488.

11 Damon M,Vincent A,Lombardi A,et al. First evidence of uncoupling protein 2(UCP-2) and 3 (UCP-3) gene expression in piglet skeletal muscle and adipose tissue[J].Gene,2000,246(1～2):133～141.

12 Fleury C, Sanchis D. The mitochondrial uncoupling protein-2: Current status[J]. Int J Biochem Cell Biol, 1999, 31(11): 1261～1278.

13 Girousse A, Avernier G, Tiraby C, et al. Transcription of the human uncoupling protein 3 gene is governed by a complex interplay between the promoter and intronic sequences[J]. Diabetologia, 2009, 52: 1638～1646.

14 Gronostajski R M. Roles of the NFI/CTF gene family in transcription and development[J]. Gene, 2000, 249: 31～45.

15 Ho P W, Ho J W, Liu H F, et al. Mitochondrial neuronal uncoupling proteins:A target for potential disease-modification in Parkinson's disease[J].Translational Neuro Degeneration,2012, 1(1):3～12.

16 Hughes J, Criscuolo F. Evolutionary history of the UCP gene family: Gene duplication and selection[J], BMC Evolutionary Biology,2008, 8(3):306～319.

17 Kim T H, Barrera L O, Qu C, et al. Direct isolation and identification of promoters in the human genome[J]. Genome Res, 2005, 15:830～839.

18 Larkin S, Mull E, Miao W, et al. Regulation of the third member of the uncoupling protein family,UCP3, by cold and thyroid hormone[J]. Biochem Biophys Res Commun, 1997, 240(1): 222～227.

19 Nadjar B E, Hinits Y, Funkenstein B, et al. Structural and functional analysis of myostatin-2 promoter alleles from the marine fish Sparus aurata: Evidence for strong muscle-specific promoter activity and post-transcriptional regulation[J]. Molecular and Cellular Endocrinology, 2012,361(1/2):51～68.

20 Nowacka-Woszuk J,Szczerbal I,Fijak-Nowak H,et al. Chromosomal localization of 13 candidate genes for human obesity in the pig genome[J].J Appl Genet,2008,49(4):373～377.

21 Ramsay T G,Mitchell A D,Richards M P.Uncoupling protein expression in skeletal muscle and adipose tissue in response to in vivo porcine somatotropin treatment[J].Domestic Animal Endocrinology,2008a,35(2):130～141.

22 Ramsay T G,Mitchell A D.Impact of dietary protein content on uncoupling protein mRNA abundance in swine[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,2008b,149(4):562～571.

23 Ricquier D, Bouillaud F. The uncoupling protein homologues: UCP1, UCP2, UCP3, StUCP and AtUCP[J]. Biochem J, 2000, 345(2): 161～179.

24 Sherman E L,Nkrumah J D,Murdoch B M,et al.Polymorphisms and haplotypes in the bovine neuropepfide Y, growth hormone receptor, ghrelin,insulin-like growth factor 2,and uncoupling proteins 2 and 3 genes and their associations with measures of growth,performance,feed eficiency,and carcass merit in beef cattle[J].Journal of Animal Science,2008,86(1):1～16.

25 Surwit R S, Wang S, Petro A E, et al. Diet-induced changes in uncoupling proteins in obesity-prone and obesity-resistant strains of mice[J]. Proc Natl Acad Sci USA, 1998, 95(7): 4061～4065.

[1]	宋兴超, 孟金柱, 赵园园, 吴震洋, 安清明. 贵州白山羊MyoG基因启动子区序列克隆及生物信息学分析[J]. 中国畜牧兽医, 2023, 50(3): 893-903.
[2]	高晓敏, 周舒简, 陈晨, 金晶, 胡菜, 张晨, 左其生, 张亚妮, 陈国宏, 李碧春. STAT1和组蛋白乙酰化修饰调控鸡lncRNA-BMP4转录研究[J]. 中国畜牧兽医, 2022, 49(9): 3321-3332.
[3]	刘华云, 张磊, 徐桂利, 张天浩, 谢遇春, 段玲欣, 刘铮铸, 巩元芳. 银黑狐MC1R基因核心启动子区的鉴定[J]. 中国畜牧兽医, 2022, 49(7): 2442-2450.
[4]	杜鑫泽, 马鑫浩, 张殿琦, 杜嘉伟, 马婧, 谢琨成, 何杰, 昝林森. bta-miR-34b/c和bta-miR-449a/b/c靶基因预测及生物信息学分析[J]. 中国畜牧兽医, 2022, 49(7): 2451-2461.
[5]	翟哲, 陈思, 吴艳茹, 王雪梅, 李崇瑞, 刘志勇, 陈巧玲, 王凤阳, 杜丽, 李昌, 金宁一. 绵羊CCL19基因启动子活性分析及其转录调控元件筛选[J]. 中国畜牧兽医, 2022, 49(4): 1213-1222.
[6]	陈哲, 杨鹏霞, 雷明明, 陈佳彬, 闫乐艳. 鹅p21基因克隆、启动子分析及转录调控研究[J]. 中国畜牧兽医, 2022, 49(2): 405-415.
[7]	李玲玉, 郑海英, 陈明棠, 唐丽萍, 尚江华, 杨春艳. 卵母细胞来源对水牛体细胞核移植胚胎发育潜能的影响[J]. 中国畜牧兽医, 2022, 49(1): 241-247.
[8]	吕巧, 邹超霞, 吴永梅, 张瑞门, 郑自华, 石德顺, 韦英明, 邓彦飞. STAT3基因对广西黄牛肌肉干细胞成肌诱导分化的调控研究[J]. 中国畜牧兽医, 2021, 48(8): 2771-2777.
[9]	刘鹏程, 张继, 邱淦远, 龚俞, 李雪松, 李维, 张依裕, 刘若余. 关岭牛TBC1D7基因多态性及其与生长性状的关联分析[J]. 中国畜牧兽医, 2021, 48(8): 2853-2863.
[10]	陆杏蓉, 段安琴, 马小娅, 梁莎莎, 邓廷贤. 水牛HSD17B1基因启动子荧光素酶报告基因载体构建与分析[J]. 中国畜牧兽医, 2021, 48(10): 3512-3521.
[11]	杨琴, 邓肖玉, 赵晓丽, 张欢, 席静, 易继海, 王勇, 张倩, 王震, 陈创夫. 布鲁氏菌Ⅳ型分泌系统对树突状细胞自噬、胞内生存和炎症反应的影响[J]. 中国畜牧兽医, 2021, 48(1): 257-264.
[12]	赵海东, 邬明丽, 陈平博, 韦燕佩, 王淑辉, 孙秀柱. IGF2基因表达调控及其遗传变异在动物生长发育中的研究进展[J]. 中国畜牧兽医, 2020, 47(6): 1844-1852.
[13]	艾锦新, 龙安炬, 罗卫星, 蔡惠芬. 贵州地方山羊BMPR-ⅠB基因启动子区多态性及生物信息学分析[J]. 中国畜牧兽医, 2020, 47(4): 1000-1008.
[14]	陈伟, 许厚强, 陈祥, 覃海, 吴雨, 黄明捷, 卢贤君. 牛UCP3和MYH1基因启动子在C2C12和3T3-L1细胞中的启动活性分析[J]. 中国畜牧兽医, 2020, 47(12): 3825-3832.
[15]	张冬杰, 汪亮, 刘洋, 刘娣. 调控民猪Tβ4基因转录因子的筛选与分析[J]. 中国畜牧兽医, 2019, 46(9): 2535-2542.