飞行时间质谱法检测水牛黑素皮质素受体4基因多态性

doi:10.16431/j.cnki.1671-7236.2015.07.026

›› 2015, Vol. 42 ›› Issue (7): 1800-1806.doi: 10.16431/j.cnki.1671-7236.2015.07.026

飞行时间质谱法检测水牛黑素皮质素受体4基因多态性

邓廷贤, 庞春英, 朱鹏, 段安琴, 陆杏蓉, 杨炳壮, 梁贤威

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

收稿日期:2015-01-20 出版日期:2015-07-20 发布日期:2015-07-23
通讯作者: 梁贤威 E-mail:liangbri@126.com
作者简介:邓廷贤(1983-),男,湖南郴州人,硕士,助理研究员,研究方向:水牛分子遗传学,E-mail:dtx282000@163.com
基金资助:
国家国际科技合作专项项目(2014DFA31970);桂渔牧科(1304512);水牛基(1404001)

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

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

Received:2015-01-20 Online:2015-07-20 Published:2015-07-23

摘要/Abstract

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

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.

Key words: buffalo; matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; MC4R gene; single nucleotide polymorphisms

中图分类号:

S813.3

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]. , 2015, 42(7): 1800-1806.

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]. , 2015, 42(7): 1800-1806.

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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

Rapid Detection of SNPs in Buffalo MC4R Gene by Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

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