美洲水貂刺鼠信号蛋白基因SNPs检测及其与毛色表型的关联分析  

Single Nucleotide Polymorphisms Detection of Agouti Gene and Its Association with Coat Color Phenotype in American Mink(Neovison vison)

作　　者：宋兴超[1] 徐超[1] 刘宗岳[1] 岳志刚[1] 丛波[1] 刘琳玲[1] 杨福合[1] 

机构地区：[1]中国农业科学院特产研究所,吉林省特种经济动物分子生物学省部共建国家重点实验室,长春130112

出　　处：《畜牧兽医学报》2016年第4期723-732,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点基础研究发展计划-973项目(2012CB722907);水貂优良种质资源及选育技术引进-948项目(2014-Z8)

摘　　要：旨在检测刺鼠信号蛋白(Agouti signaling protein,Agouti)基因外显子2、内含子2、外显子3及部分内含子3的单核苷酸多态性(Single nucleotide polymorphisms,SNPs)在不同毛色美洲水貂群体中的分布,探讨该基因变异与水貂被毛颜色表型的相关性。以金州黑水貂、吉林白水貂、银蓝水貂、咖啡水貂和珍珠色水貂共计430个样本的血液基因组DNA为模板,采用PCR扩增和Sanger双脱氧链终止测序技术,对Agouti基因序列进行SNPs检测,并将突变位点与水貂毛色表型进行关联分析。结果表明,获得的美洲水貂Agouti基因长度为2 510bp,内含子2存在4个SNPs:g.18G>A、g.159A>G、g.235G>T和g.1189C>T,部分内含子3检测到6个SNPs:g.252C>T、g.290A>C、g.298G>C、g.340A>G、g.343T>C和g.379T>C,在外显子2和3区域并未检测到SNPs位点。关联分析表明,Agouti基因7个SNPs(内含子2:g.1189C>T;内含子3:g.252C>T、g.290A>C、g.298G>C、g.340A>G、g.343T>C和g.379T>C)位点的基因型均与水貂毛色表型极显著相关(P<0.000 1),且部分内含子3中的5个SNPs位点(g.290A>C、g.298G>C、g.340A>G、g.343T>C和g.379T>C)可能处于紧密连锁状态。研究结果初步表明,Agouti基因可能是影响美洲水貂被毛颜色的候选基因或与决定毛色性状主效基因相连锁的分子标记。The aim of this study was to research single nucleotide polymorphisms of agouti signal protein（Agouti）gene and analyze the association of SNPs with coat color phenotype in American mink（Neovison vison）. The DNA of blood from 5 mink breeds（Jinzhou black,Jilin white, Sil- verblue,Coffee and Pearl mink）that possessed significant difference in coat color phenotype were selected as templates,SNP sites were screened by PCR and Sanger double chain termination se- quencing technology, and then the relationship between the mutation sites of Agouti gene and coat color phenotype were analyzed in 430 minks. The results showed that the obtained mink Agouti gene was 2 510 bp in length. Total 10 SNPs were screened from 430 individuals of 5 coat color breeds,of which 4 SNPs （g. 18G〉A,g. 159A〉G,g. 235G〉T and g. 1189C〉T）were located in intron 2 and 6 SNPs（g. 252C〉T,g. 290A〉C,g. 298G〉C,g. 340A〉G,g. 343T〉C and g. 379T〉C） were detected from partial intron 3 of Agouti gene,while no SNPs was discovered in exon 2 and 3. Association analysis of 7 SNPs in Agouti gene with coat color phenotype indicated that all sites were very significantly（P〈0. 000 1）correlated with coat color phenotype of American mink. In addition,the 5 loci including g. 290A〉C,g. 298G〉C,g. 340A〉G,g. 343T〉C and g. 379T〉C might show closely linkaged phenomenon. Results of the present study indicated that Agouti gene might be the candidate gene or linked with the major gene affecting the coat color phenotype of American mink.

关 键 词：美洲水貂 Agouti基因 毛色表型 单核苷酸多态性 关联分析 

分 类 号：S865.222[农业科学—野生动物驯养]