地方黄牛MyoD1基因启动子多态性分析及其相关转录因子SF1验证

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摘要摘要骨骼肌的形成影响着动物的生长发育，而成肌因子的调控，将决定动物的生长性能高低，是极其重要的经济指标之一。为筛选不同品种黄牛(Bos taurus)成肌细胞决定基因1(myogenic differentiation 1, MyoD1)启动子区SNPs (single nucleotide polymorphisms)，旨在为确定与黄牛生长性状相关的分子遗传标记提供研究基础，为MyoD1基因启动子区功能元件的揭示提供依据。实验以200头贵州地方黄牛关岭牛、思南牛、黎平牛、威宁牛(各50头)为研究对象，分析MyoD1基因的单核苷酸突变位点。在4个实验群体中共发现5个SNPs，其中关岭牛、思南牛、黎平牛的突变位点相同，都为g.-1141T>C、g.-871C>T；威宁牛的突变位点有4个，分别为g.-1778T>C、g.-871C>T、g.-797T>C、g.-761T>C。关联性分析结果表明，关岭牛MyoD1基因启动子g.-1141T>C、g.-871C>T位点分别对腹围、体斜长和胸围有显著影响，黎平牛MyoD1基因启动子g.-1141T>C、g.-871C>T两个位点均对管围存在显著影响，思南牛MyoD1基因启动子的g.-1141T>C位点对坐骨端宽有显著影响(P＜0.05)，而g.-871C>T位点对生长指标没有显著性影响(P＞0.05)。威宁牛MyoD1基因启动子g.-1778T>C、g.-871C>T、g.-797T>C、g.-61T>C位点分别对体斜长、腰高和坐骨端宽有显著影响(P＜0.05)。采用酵母单杂交实验对MyoD1基因启动子和转录因子SF1基因验证，结果发现SF1基因确能与MyoD1基因启动子结合的转录因子。利用双荧光素酶报告系统检测到SF1基因对MyoD1基因启动子相对荧光素酶活性升高。当SF1基因转录位点产生g.-1141T>C突变，为CC基因型时，对3个黄牛品种(关岭牛, 黎平牛, 思南牛)的腹围、管围、坐骨端宽等生长指标影响显著，SF1对MyoD1基因的启动活性及调控功能有重要影响。

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	周迪
	许厚强
	陈伟
	赵佳福
	段志强

关键词 ： MyoD1基因, 启动子, SF1转录因子, 酵母单杂交

Abstract：Abstract The formation of skeletal muscle affects the growth and development of animals. Through screening SNPs of MyoD1 gene promoter in different breeds of cattle (Bos taurus), and studying its effect on growth body size traits and promoter function, to further provide research basis for the molecular genetic markers of growth related traits and MyoD1 gene promoter revealed promoter functional components. 200 Guizhou native cattle (including Guanling cattle, Sinan cattle, Liping cattle, Weining cattle, 50 heads , respectively) as the research material, the experiment amplified MyoD1 promoter and used direct sequencing method to detect SNPs. The experiment found 5 SNPs in four groups, g.-1141T>C、g.-871C>T in Guanling cattle, Sinan cattle, Liping cattle and g.-1778T>C、g.-871C>T、g.-797T>C、g.-761T>C in Weining cattle; Correlation analysis results showed that in Guanling cattle -1141 T>C, g.-871C>T sites had significant influence on abdominal extension girth, body length and chest girth(P＜0.05), respectively; in Liping cattle -1141T>C, g.-871C>T sites had significant influence on cannon circumference(P＜0.05); in Sinan cattle g.-1141T>C site had significant influence on hucklebone width, while g.-871C>T had no significant influence on growth traits; in Weining cattle g.-1778T>C、g.-871C>T、g.-797T>C、g.-761T>C sites had significant influence on body length, hucklebone width and wasitband height(P＜0.05). Using the yeast one-hybrid (Y1H) system, the results found the SF1 transcription factor could be combined with the MyoD1 promoter; using dual luciferase report system, SF1 transcription factor increased the MyoD1 promoter activity. When the SF1 transcription site produced g.-1141T>C mutation, it had significant effect on abdominal girth, cannon circumference and hucklebone width and so on growth traits in three cattle(Guanling cattle, Liping cattle, Sinan cattle). SF1 has important influence for the activation and regulation of the MyoD1 gene.

Key words： MyoD1gene Promoter SF1 transcription factor Yeast single-hybridization experiment

收稿日期: 2017-09-05 出版日期: 2018-05-02

基金资助:国家自然科学基金

通讯作者: 许厚强 E-mail: houqiang0524@yahoo.com;gzdxxhq@163.com

引用本文:

周迪许厚强陈伟赵佳福段志强. 地方黄牛MyoD1基因启动子多态性分析及其相关转录因子SF1验证[J]. , 2018, 26(5): 743-755.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2018/V26/I5/743

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