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飞行时间质谱法检测水牛黑素皮质素受体4基因多态性
引用本文:邓廷贤,庞春英,朱鹏,段安琴,陆杏蓉,杨炳壮,梁贤威.飞行时间质谱法检测水牛黑素皮质素受体4基因多态性[J].中国畜牧兽医,2015,42(7):1800-1806.
作者姓名:邓廷贤  庞春英  朱鹏  段安琴  陆杏蓉  杨炳壮  梁贤威
作者单位:中国农业科学院广西水牛研究所, 农业部(广西)水牛遗传繁育重点实验室, 南宁 530001
基金项目:国家国际科技合作专项项目(2014DFA31970);桂渔牧科(1304512);水牛基(1404001)
摘    要:应用基质辅助激光解吸电离飞行时间质谱技术(MALDI-TOF-MS)检测水牛黑素皮质素受体4(MC4R)基因的多态性(SNPs),为今后建立水牛辅助标记选择策略奠定基础。以水牛MC4R基因序列为模板,利用PCR扩增测序法筛查了水牛MC4R基因13个SNPs。为进一步验证筛选的可靠性,应用MALDI-TOF-MS针对380头水牛检测了8个标记的基因型。结果显示,平均检出率为98.0%,能够准确的检测出3种常见的基因型。所检SNP位点的平均最小等位基因频率(MAF)为0.24,平均杂合度为0.28,平均多态信息含量(PIC)为0.23。本研究建立的基于PCR测序法联合MALDI-TOF-MS检测水牛MC4R基因多态性的方法,具有快速、可靠和准确的特点,为今后开展水牛生产性状基因的SNP分型和检测等研究奠定了基础。

关 键 词:水牛  飞行时间质谱  黑素皮质素受体4基因  SNPs  
收稿时间:2015-01-20

Rapid Detection of SNPs in Buffalo MC4R Gene by Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry
DENG Ting-xian,PANG Chun-ying,ZHU Peng,DUAN An-qin,LU Xing-rong,YANG Bing-zhuang,LIANG Xian-wei.Rapid Detection of SNPs in Buffalo MC4R Gene by Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry[J].China Animal Husbandry & Veterinary Medicine,2015,42(7):1800-1806.
Authors:DENG Ting-xian  PANG Chun-ying  ZHU Peng  DUAN An-qin  LU Xing-rong  YANG Bing-zhuang  LIANG Xian-wei
Institution:Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Ministry of Agriculture (Guangxi), Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning 530001, China
Abstract:The study used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for rapid determination of single nucleotide polymorphisms (SNPs) in buffalo MC4R gene,which provided a foundation for constructing the marker-assisted selection(MAS)strategy.The cDNA sequence of buffalo MC4R gene was screened rapidly based on PCR amplification and sequencing method,and in which 13 SNPs were found.To further identify the reliability of screening system,8 SNPs were genotyped in 380 buffaloes using the MALDI-TOF-MS technology.The results revealed that 8 SNPs had an average of 98.0% in call rate,and in which three genotypes could be accurately distinguished.The average minor allele frequency (MAF),average heterozygosity and polymorphism information content (PIC) value of the 8 markers was 0.24,0.28 and 0.23,respectively.Our findings indicated that the rapid determination method of MALDI-TOF-MS with PCR and sequencing with highly accurate properties was a useful tool for identifying the SNPs of buffalo MC4R gene that laid the foundations for studying SNP genotype in buffalo genes associated production traits.
Keywords:buffalo  matrix-assisted laser desorption/ionization time-of-flight mass spectrometry  MC4R gene  single nucleotide polymorphisms  
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