畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (9): 1824-1829.doi: 10.11843/j.issn.0366-6964.2016.09.010

• 遗传繁育 • 上一篇    下一篇

不同类型牛GPR54基因启动子变异与其表达水平的关联性

王学故1,邓丹妹1,程晋1,王扬1,李彤1,陈宏权 1,2*   

  1. (1.安徽农业大学动物科技学院,合肥 230036;  2.安徽畜禽遗传资源保护与生物育种省级实验室,合肥230036)
  • 收稿日期:2016-03-07 出版日期:2016-09-23 发布日期:2016-09-23
  • 通讯作者: 陈宏权,教授,博导,主要从事动物分子遗传育种研究,E-mail:chqchq@ahau.edu.cn
  • 作者简介:王学故(1991-),男,安徽利辛人,硕士生,主要从事动物分子遗传育种研究,E-mail:1055069260@qq.com
  • 基金资助:

    国家自然科学基金项目(31272402)

The Relationship between the Expression Variations of GPR54 Gene and the Polymorphisms of GPR54 Promoters in Different Types of Bovines

WANG Xue-gu 1,DENG Dan-mei 1,CHENG Jin 1,WANG Yang 1,LI Tong 1,CHEN Hong-quan 1,2*   

  1. (1.College of Animal Science and Technology, Anhui Agricultural University,Hefei 230036,China;2.Key Laboratory of Anhui Local Livestock and Poultry Genetic Resources Conservation and Biobreeding of Anhui Province,Hefei 230036,China)
  • Received:2016-03-07 Online:2016-09-23 Published:2016-09-23

摘要:

旨在研究不同类型牛GPR54基因启动子的变异对其表达水平的影响,以揭示牛GPR54基因多态性与牛早熟性之间的关联性。提取荷斯坦牛、皖东黄牛及江淮水牛3种牛血液白细胞总RNA,通过PCR、测序、RT-PCR和双荧光素酶报告基因的方法,检测不同牛GPR54基因启动子区的多态性,启动子效率以及GPR54基因的表达水平,分析启动子变异在牛品种、年龄和性别间对GPR54基因表达的效应。结果表明,GPR54基因启动子存在多个SNP位点,3种牛的启动子型存在明显的差异,其中荷斯坦牛GPR54基因启动子效率是江淮水牛的4.44倍,是皖东黄牛的1.99倍,而皖东黄牛启动子效率是江淮水牛的2.24倍。RT-PCR显示,青年牛GPR54基因表达水平高于成年牛;荷斯坦牛高于皖东黄牛和水牛,水牛的GPR54表达水平最低;母牛的GPR54基因表达水平极显著高于公牛(P<0.01)。综上表明,不同牛的早熟性与GPR54启动子存在密切的关联性。

Abstract:

 The aim of the study is to investigate the impact of the variations of GPR54 promoter on the expression levels of GPR54 gene in Holstein (HT),Wandong cattle (WD) and Jianghuai buffalo (JH),therefore further unveil the association between the polymorphisms of GPR54 gene promoter and the sexual precocity.Total RNAs in leukocytes of HT,WD and JH were extracted and the polymorphism of the promoter region were detected by PCR and sequencing.Furthermore,the efficiency of the promoters were determined via RT-PCR and luciferase expression from the fusion reporter gene.Various SNPs were detected in the promoter regions of GPR54 genes in HT,WD and JH with significant difference.Moreover,it was shown that the promoter efficiency of HT was 4.44 times and 1.99 times higher compared to JH buffalo and WD respectively.Meanwhile the promoter efficiency of WD was 2.24 times higher than JH.Notably,the RT-PCR results showed that GPR54 gene expression levels in youngsters were dramatically higher than that of the adults especially in HT than in WD and JH.JH showed the weakest expression level of GPR54 while the cows exhibited the strongest GPR54 expression level compared to ox (P<0.01).These results indicated that the polymorphisms of GPR54 promoter and the bovine sexual precocity were evidently correlated.

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