实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (4): 363-370.DOI: 10.12300/j.issn.1674-5817.2023.055

• 华东会议优秀论文展 • 上一篇    下一篇

BALB/cA.Cg.SHJH hr 小鼠的培育及其相关遗传学特性分析

谈小倩1, 杨颢2, 唐慧青1, 瞿伟1, 李亮1, 钱珍1, 顾坚忠1, 徐平1()(), 肖君华2()()   

  1. 1.上海吉辉实验动物饲养有限公司, 上海 201611
    2.东华大学生物科学与技术研究所, 上海 201620
  • 收稿日期:2023-04-28 修回日期:2023-06-08 出版日期:2023-08-25 发布日期:2023-09-14
  • 通讯作者: 徐 平(1964—),男,研究员,研究方向为实验动物资源和低温生物学。E-mail: pingxu@jh-labanimal.com。ORCID: 0000-0001-6744-8908;
    肖君华(1968—),男,教授,研究方向为小鼠遗传学。E-mail: xiaojunhua@dhu.edu.cn。ORCID:0000-0002-1977-372X
  • 作者简介:谈小倩(1982—),女,执业兽医师,研究方向为实验动物模型和实验动物疾病。E-mail: xiaoqian_tan@163.com
  • 基金资助:
    上海市科技创新行动计划项目“自发性早衰小鼠的种群建立及相关机制研究”(21140909100)

Creation and Analysis of Related Genetic Characteristics of BALB/cA.Cg.SHJH hr Mice

Xiaoqian TAN1, Hao YANG2, Huiqing TANG1, Wei QU1, Liang LI1, Zhen QIAN1, Jianzhong GU1, Ping XU1()(), Junhua XIAO2()()   

  1. 1.Shanghai Jihui Laboratory Animal Care Co. , Ltd. , Shanghai 201611, China
    2.Institute of Biological Science and Technology, Donghua University, Shanghai 201620, China
  • Received:2023-04-28 Revised:2023-06-08 Published:2023-08-25 Online:2023-09-14
  • Contact: XU Ping (ORCID: 0000-0001-6744-8908), E-mail: pingxu@jh-labanimal.com;
    XIAO Junhua (ORCID: 0000-0002-1977-372X), E-mail: xiaojunhua@dhu.edu.cn;

摘要:

目的 将自发突变培育的SHJH hr 小鼠的突变Hr基因导入到遗传背景清晰的BALB/cAShjh近交系小鼠,通过突变基因筛查和遗传学特性分析为后续研究Hr基因突变导致异常表型的分子机制以及该模型的推广应用提供依据。 方法 采用在表型监测指导下回交-互交的育种方法,将自发突变培育的SHJH hr 小鼠的突变基因以同源突变导入方式导入到近交系BALB/cAShjh小鼠,经10代后培育成BALB/cA.Cg.SHJH hr (简称C.Cg.SHJH hr )小鼠。通过多重PCR建库后二代测序的方法,分析C.Cg.SHJH hr 小鼠中随机分布于基因组上的90个单核苷酸多态性(single nucleotide polymorphism,SNP)检测位点基因型。然后按照GB/T 14927.1—2008的方法,对C.Cg.SHJH hr 小鼠中14个生化位点标记基因进行检测。最后利用全基因组外显子测序技术,检测该小鼠的突变基因。 结果 自2018年5月至2022年3月,共进行了10代的回交-互交,完成了C.Cg.SHJH hr 小鼠品系的构建。在检测的90个SNP位点中,除了rs13484115、rs13484116两个位点不同外,C.Cg.SHJH hr 小鼠的其他位点基因型均与受体小鼠BALB/cAShjh相同;生化位点标记基因检测结果显示,C.Cg.SHJH hr 小鼠的14个位点全部与受体小鼠相同;全基因组外显子测序发现,该小鼠相比受体小鼠品系存在109个位点突变,包括同义突变71个,终止密码子增加1个,错义突变37个,涉及蛋白质序列改变的基因20个(包括已报告的Hr基因)。 结论 成功培育了突变导入系C.Cg.SHJH hr 小鼠,通过全基因组外显子测序分析发现了3个主要引起表型变异的Hr突变基因及关联突变基因,为该小鼠在生物医学研究中的拓展应用提供了基础数据。

关键词: BALB/cA.Cg.SHJH hr 小鼠, 外显子测序, 突变基因, 遗传学特性

Abstract:

Objective To introduce the Hr gene of spontaneously mutated SHJH hr mice into BALB/cAShjh inbred mice with clear genetic background,and provide a basis for study on the molecular mechanism of Hr gene mutation-induced abnormal phenotype and the application of this model. Methods Using a backcross-intercross breeding method guided by phenotypic monitoring, mutant genes from SHJH hr mice bred by spontaneous mutation were introduced into inbred BALB/cAShjh mice by homozygous mutation introgression, and the mice were bred into BALB/cA.Cg.SHJH hr (abbreviated as C.Cg.SHJH hr ) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJH hr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJH hr mice according to the method of GB/T 14927.1-2008. Finally, whole genome exon sequencing was utilized to detect the mutated genes in this mouse. Results From May 2018 to March 2022, a total of 10 generations of backcross-intercross were conducted to complete the construction of the C.Cg.SHJH hr mouse line. Among the 90 SNPs loci detected, except for rs13484115 and rs13484116, all the other loci had the same genotype as the recipient mice BALB/cAShjh. The results of biochemical marker gene detection showed that all the 14 loci of the mouse were the same as those of the recipient mouse. Whole genome exon sequencing found that the mouse had 109 site mutations compared with the recipient mouse strain, including 71 synonymous mutations, 1 stopgain, 37 missense mutations, and 20 genes involved in protein sequence alterations (including the reported Hr gene). Conclusion C.Cg.SHJH hr mice were created. Through exon sequencing and genetic analysis, three Hr mutated genes and associated mutated genes that mainly cause phenotypic variations were identified, which provides a basis for expanding the application of C.Cg.SHJH hr mice in biomedical research.

Key words: BALB/cA.Cg.SHJH hr mice, Exon sequencing, Mutation, Genetic characteristics

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