DRD1基因5'侧翼区多态性及其与公鸡鸡冠性状的相关性  

Association of DRD1 Gene Polymorphism in the 5' Regulatory Region with the Cock's(Gallus gallus) Comb Traits

作　　者：李春苗[1] 黎寿丰[1] 赵振华[1] 黄华云[1] 王钱保[1] 薛龙岗[1] 

机构地区：[1]中国农业科学院家禽研究所,扬州225125

出　　处：《农业生物技术学报》2017年第4期526-536,共11页Journal of Agricultural Biotechnology

基　　金：现代农业产业技术体系建设专项(No.CARS-42-Z06);江苏省科技支撑项目(No.BE2013386);江苏省科技支撑项目(No.BE2014364);扬州市自然科学基金项目(No.YZ2015104);扬州市现代农业项目(No.YZ2015041)

摘　　要：多巴胺D1受体(dopamine receptor D1,DRD1)能促进或抑制鸟类大脑中催乳素(prolactin,PRL)的分泌,在鸡(Gallus gallus)的下丘脑和垂体高表达,并与生殖系统相关联。为探讨DRD1基因5'调控区多态性与鸡早熟性状的关系,本研究以中国农业科学院家禽研究所选育的140只W系公鸡为实验材料。利用限制性片段长度多态性聚合酶反应技术(cleaved amplification polymorphism sequence-tagged sites,CAPs or PCR-RFLP)检测DRD1基因的SNPs,并分析其多态性与公鸡77、84和91日龄的体重、冠高1、冠高2和冠长之间的关联性。结果显示,在距CDS-570位点发现1个突变位点(G→A),G-570A位点检测到GG、GA和AA 3种基因型。关联性分析显示,77日龄时,GG型个体体重显著高于AA与AG型个体(P<0.05),不同基因型个体冠高1、冠高2和冠长差异均不显著;84日龄时,不同基因型个体间体重、冠高1、冠高2和冠长差异均不显著;91日龄时,不同基因型个体冠高1和冠长差异均显著,表现为GG型个体冠高1显著高于AA与AG型个体(P<0.05);GG型个体冠长显著长于AA与AG型个体(P<0.05)。构建鸡DRD1基因5'调控区DRD1/883序列不同基因型双荧光素酶报告基因表达载体(p GL3-DRD1/883/promoter),脂质体转染法转染至293T细胞,检测不同基因型的启动子活性,结果显示,DRD1/883G作为启动子时,萤火虫对海肾荧光素酶的强度比值要比使用DRD1/883A启动子效率提高13.3倍,呈高效表达(P<0.01)。由此可见,对于鸡早熟性状而言,GG是最有利基因型,携带DRD1/883G个体具有较高的转录水平,说明该基因可能对鸡早熟性状有一定的调控作用,该研究结果为DRD1基因的表达和功能研究及优质肉鸡的选育提供理论依据。Dopamine receptor D1（DRD1） can promote or inhibit the secretion of prolactin in the birds＇ brain. It is associated with the reproductive system and highly expressed in the hypothalamus and pituitary. The experiment was designed to study of polymorphism of 5＇ flanking regulating of DRD1 gene on sexual maturity traits in cock （Gallus gallus）, 140 W lines breeding from Poultry Institute, Chinese Academy of Agricultural Sciences were used in the experiment. The SNPs was detected by PCR-RFLP （cleaved amplification polymorphism sequence-tagged sites） and analyzed the association of polymorphisms sites with body weight, comb height 1, comb height 2 and comb length of cock at 77, 84 and 91 days of age. The results indicated that one mutation site was detected in 5＇ flanking regulating of DRD1 gene. Three genotypes （AA, AG and GG） were found at G-570A mutation locus at the distance of CDS-570 locus in 140 cocks by digesting the PCR product with MspⅠ. The relationship between DRD1 gene and the sexual maturity traits was analyzed by least squares linear model. The individuals with GG genotype had significant effect on the body weight （P〈0.05）, and had no significant effect on the comb height 1, comb height 2 and comb length of cock at 77 days of age. At 84 days of age, the individuals with different genotype of body weight comb height 1, comb height 2 and comb length were not significant. The individuals with GG genotype had significant effect on comb height 1 and comb length （P〈0.05）. The DRD1/883 promoter of DRD1 gene was amplified using PCR technology, inserted into the dual-luciferase reporter vector （pGL3-Basic） to construct pGL3-DRD1/883/promoter. 293T cell were co-transfected with a plasmid containing the pGL3-DRD1/883/promoter, pGL3-Basic and PRL-CMV, respectively. The activity of luciferase gene in 293T cell was testing using the Dual-Luciferase Reporter Assay kit. The results showed that the intensity ratio of firefly to sea renal luciferase of DRD1/883G prompter enhance 13.

关 键 词：多巴胺D1受体基因(DRD1) 多态性 性早熟性状 

分 类 号：S831.2[农业科学—畜牧学]