用于四种常用品系大鼠鉴定的单核苷酸多态性标记筛查和应用研究
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(1. 华南理工大学生物科学与工程学院,广州 510006; 2. 广东省实验动物监测所,广州 510663)

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R-33

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Screening of single nucleotide polymorphism markers and their application to identification of four common strains of rats
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(1. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China. 2. Guangdong Laboratory Animals Monitoring Institute, Guangzhou 510663)

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    摘要:

    目的 筛选出能快速鉴定四种常用品系大鼠[Wistar 大鼠?Goto-Kakizaki(GK)大鼠?Brown-Norway (BN)大鼠?Sprague-Dawley(SD)大鼠]的单核苷酸多态性(single nucleotide polymorphism,SNP)位点并验证其可靠性?方法 首先对Wistar 大鼠和GK 大鼠进行全基因组重测序,利用生物信息学技术将重测序结果比对到BN 大鼠参考基因组,筛查到Wistar 大鼠和GK 大鼠基因组范围内的特异性SNP,并从中进一步筛选出一组候选SNP 位点,再利用一代测序和DNA 混合池法对以上候选SNP 位点在Wistar?GK?BN?SD 四种大鼠群体中进一步筛查与验证?结果 Wistar 和GK 大鼠通过比对分别得到94 800 和106 019 个纯合特异性SNP,其中有56 216 个SNP 为Wistar 和GK 大鼠所共有,且发现BN 大鼠参考基因组?Wistar 大鼠和GK 大鼠三者基因型互不相同的位点仅有38个,从中挑选了15 个作为候选SNP 位点,最终在Wistar?GK?BN?SD 四种大鼠群体验证中得到13 个可以区分四个品系大鼠的SNP 标记及其在各个品系中的基因型?结论 筛查并验证了能区分Wistar?GK?BN 和SD 四种大鼠的13 个SNP 标记,这些SNP 标记可快速区分出Wistar?GK?BN 和SD 大鼠?

    Abstract:

    Objective To screen out single nucleotide polymorphisms (SNPs) that can be used to rapidly identify four common strains (Wistar, GK, BN, and SD) of rats and verify their reliability. Methods First, whole genome resequencing was performed on Wistar and GK rats. Using bioinformatic techniques to align the resequencing reads to the BN rat reference genome to detect specific SNPs within the genome of Wistar and GK rats, we selected a group of candidate SNPs. Then, we used Sanger sequencing and the DNA pooling method to screen and validate the candidate SNPs in four strains of rats. Results 94 800 and 106 019 homozygous-specific SNPs were detected in the Wistar and GK rats,respectively, of which 56 216 were shared by Wistar and GK rats. Only 38 SNPs were found to show different genotypes among BN, Wistar, and GK rat reference genomes, of which 15 were selected as candidate SNPs. Finally, 13 SNPs were screened, and the genotype of each locus in the four strains of rats was identified. Conclusions Thirteen SNP markers were identified and validated. These SNP markers can be used to rapidly distinguish Wistar, GK, BN, and SD rats.

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曾玉琳,蒙裕欢,张钰,杜红丽.用于四种常用品系大鼠鉴定的单核苷酸多态性标记筛查和应用研究[J].中国比较医学杂志,2018,28(11):36~41.

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  • 收稿日期:2018-02-26
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  • 在线发布日期: 2018-12-18
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