Analysis of Microsatellite Polymorphism in Inbred Knockout MiceReport as inadecuate

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Previously, we found that the genotype of 42 out of 198 mouse microsatellite loci, which are distributed among all chromosomes except the Y chromosome, changed from monomorphism to polymorphism CMP in a genetically modified inbred mouse strain. In this study, we further examined whether CMP also relates to the homologous recombination in gene knockout KO mouse strains. The same 42 microsatellite loci were analyzed by polymerase chain reaction PCR in 29 KO inbred mouse strains via short tandem sequence repeat STR scanning and direct sequence cloning to justify microsatellite polymorphisms. The C57BL-6J and 129 mouse strains, from which these 29 KO mice were derived, were chosen as the background controls. The results indicated that 10 out of 42 23.8% loci showed CMP in some of these mouse strains. Except for the trinucleotide repeat locus of D3Mit22, which had microsatellite CMP in strain number 9, the core sequences of the remaining 41 loci were dinucleotide repeats, and 9 out of 41 21.95% showed CMPs among detected mouse strains. However, 11 out of 29 37.9% KO mice strains were recognized as having CMPs. The popular dinucleotide motifs in CMP were TGn 50%, 2-4, followed by GTn 27.27%, 3-11 and CAn 23.08%, 3-13. The microsatellite CMP in CTn and AGn repeats were 20% 1-5. According to cloning sequencing results, 6 KO mouse strains showed insertions of nucleotides whereas 1 showed a deletion. Furthermore, 2 loci D13Mit3 and D14Mit102 revealed CMP in 2 strains, and mouse strain number 9 showed CMPs in two loci D3Mit22 and D13Mit3 simultaneously. Collectively, these results indicated that microsatellite polymorphisms were present in the examined inbred KO mice.

Author: Baofen Zuo , Xiaoyan Du , Jing Zhao, Huixin Yang, Chao Wang , Yanhua Wu, Jing Lu, Ying Wang, Zhenwen Chen



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