实验动物与比较医学 ›› 2016, Vol. 36 ›› Issue (1): 24-31.DOI: 10.3969/j.issn.1674-5817.2016.01.005

• 论著 • 上一篇    下一篇

单核苷酸多态性(SNP)在近交系小鼠遗传检测中的应用

赵丽亚1,2, 张蓉1,2, 赵莹1,2, 邢正弘1,2, 陈国强1,2   

  1. 1.上海西普尔-必凯实验动物有限公司, 上海 201203;
    2.上海实验动物研究中心, 上海 201203
  • 收稿日期:2015-06-29 出版日期:2016-02-25 发布日期:2016-02-25
  • 作者简介:赵丽亚(1981-), 女, 硕士。研究方向: 分子遗传 学。E-mail: zhaoliya3002@163.com
  • 基金资助:
    上海市科学技术委员会科研计划项目(12140900400)

Application of Single Nucleotide Polymorphism Genotyping Panel in Genetic Monitoring of Laboratory Mice

ZHAO Li-ya1, ZHANG Rong1, ZHAO Ying1, XING Zheng-hong1, CHEN Guo-qiang1   

  1. 1. Sino-British SIPPR/BK Laboratory Animal Co., Ltd., Shanghai 201203, China;
    2. Shanghai Laboratory Animal Research Center, Shanghai 201203, China
  • Received:2015-06-29 Online:2016-02-25 Published:2016-02-25

摘要: 目的 利用单核苷酸多态性(SNP)位点构建多重聚合酶链反应和链接酶反应(PCR-LDR)方案,为近交系小鼠的遗传检测提供一种快速、简便的方法。方法 在SNP公共数据库中挑选51个SNP位点,该51个位点分布于每条常染色体和性染色体,共分为5组,进行多重PCR-LDR方案构建,并将该方案作为遗传质量检测方法对采集的7个近交系样本进行检测。结果 在送检的7个近交系小鼠样本中,纯合度都为100%,相同来源相同品系小鼠的遗传背景一致,但在不同单位的近交系小鼠中,某些SNP位点有差异,CBA/Ca/BKl和CBA/JSlac在a9、a10、b5、b9、c1、c12、e1、e2位点的遗传检测结果不同,C57BL/6/BKl和C57BL/6JSlac在c7、c10位点的遗传检测结果不同。另外,两家公司各有5个位点在所有品系中基因型相同,因此有效位点数各为46个。结论 构建的多重PCR-LDR方案能有效对两家动物生产单位的近交系小鼠进行检测,可用于常见近交系小鼠快速、高通量的基因分型,有利于大规模遗传质量检测和品系鉴定。

关键词: 遗传质量检测, 近交系小鼠, 单核苷酸多态性(SNP), 多重聚合酶链反应和链接酶反应(PCR-LDR), 分型方案

Abstract: Objective To develop multiple PCR-LDR genotyping protocols for monitoring genetic contamination in inbreed mice in a fast and reliable way. Method In total 51 single nucleotide polymorphisms (SNPs) from public SNP database were chosen and divided into five groups. These 51 SNPs are wildly distributed and there are least one SNP on each autosomal chromosome and the sex chromosome. A multiple PCR-LDR genotyping protocol was established as a genetic quality control approach to monitor the genotypes of 7 inbred mice. Results The homozygosity is 100% for the 7 tested mice. The genetic background of the inbred mice from the same source is identical. For the same inbred mice from different companies, several SNPs are different. The SNPs are different at a9、a10、b5、b9、c1、c12、e1、e2 for CBA/Ca/Bkl and CBA/JSlac, while the SNPs are different at c7, c10 for C57BL/6/BKl and C57BL/6JSlac. However, there are five loci which are the same in the 7 inbred mice of different companies. So there are 46 effective loci which are different in the 7 inbred mice of different companies. Conclusion The multiple PCR-LDR genotyping protocol is used to monitor the genetic differences of inbred mice from two companies. The results of this study provide a rapid and high-throughput genotyping approach, which is sufficient for genetic contamination monitoring and strain identification.

Key words: Genetic quality monitoring, Inbred mice, Single Nucleotide Polymorphisms (SNP), multiple PCR-LDR, Genotyping protocol

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