Shiyan dongwu yu bijiao yixue (Aug 2023)

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

  • TAN Xiaoqian,
  • YANG Hao,
  • TANG Huiqing,
  • QU Wei,
  • LI Liang,
  • QIAN Zhen,
  • GU Jianzhong,
  • XIAO Junhua,
  • XU Ping

DOI
https://doi.org/10.12300/j.issn.1674-5817.2023.055
Journal volume & issue
Vol. 43, no. 4
pp. 363 – 370

Abstract

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Objective To introduce the Hr gene of spontaneously mutated SHJHhr 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 SHJHhr 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.SHJHhr (abbreviated as C.Cg.SHJHhr) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJHhr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJHhr 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.SHJHhr 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.SHJHhr 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.SHJHhr mice in biomedical research.

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