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Zygosity Determination in Hairless Mice by PCR Based on Hr(hr) Gene Analysis
We analyzed the Hr gene of a hairless mouse strain of unknown origin (HR strain, http://animal.nibio.go.jp/e_hr.html) to determine whether the strain shares a mutation with other hairless strains, such as HRS/J and Skh:HR-1, both of which have an Hr(hr) allele. Using PCR with multiple pairs of prime...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Japanese Association for Laboratory Animal Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4160947/ https://www.ncbi.nlm.nih.gov/pubmed/23903062 http://dx.doi.org/10.1538/expanim.62.267 |
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author | Suzuki, Osamu Koura, Minako Noguchi, Yoko Uchio-Yamada, Kozue Matsuda, Junichiro |
author_facet | Suzuki, Osamu Koura, Minako Noguchi, Yoko Uchio-Yamada, Kozue Matsuda, Junichiro |
author_sort | Suzuki, Osamu |
collection | PubMed |
description | We analyzed the Hr gene of a hairless mouse strain of unknown origin (HR strain, http://animal.nibio.go.jp/e_hr.html) to determine whether the strain shares a mutation with other hairless strains, such as HRS/J and Skh:HR-1, both of which have an Hr(hr) allele. Using PCR with multiple pairs of primers designed to amplify multiple overlapping regions covering the entire Hr gene, we found an insertion mutation in intron 6 of mutant Hr genes in HR mice. The DNA sequence flanking the mutation indicated that the mutation in HR mice was the same as that of Hr(hr) in the HRS/J strain. Based on the sequence, we developed a genotyping method using PCR to determine zygosities. Three primers were designed: S776 (GGTCTCGCTGGTCCTTGA), S607 (TCTGGAACCAGAGTGACAGACAGCTA), and R850 (TGGGCCACCATGGCCAGATTTAACACA). The S776 and R850 primers detected the Hr(hr) allele (275-bp amplicon), and S607 and R850 identified the wild-type Hr allele (244-bp amplicon). Applying PCR using these three primers, we confirmed that it is possible to differentiate among homozygous Hr(hr) (longer amplicons only), homozygous wild-type Hr(shorter amplicons only), and heterozygous (both amplicons) in HR and Hos:HR-1 mice. Our genomic analysis indicated that the HR, HRS/J, and Hos:HR-1 strains, and possibly Skh:HR-1 (an ancestor of Hos:HR-1) strain share the same Hr(hr) gene mutation. Our genotyping method will facilitate further research using hairless mice, and especially immature mice, because pups can be genotyped before their phenotype (hair coat loss) appears at about 2 weeks of age. |
format | Online Article Text |
id | pubmed-4160947 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Japanese Association for Laboratory Animal Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-41609472014-10-21 Zygosity Determination in Hairless Mice by PCR Based on Hr(hr) Gene Analysis Suzuki, Osamu Koura, Minako Noguchi, Yoko Uchio-Yamada, Kozue Matsuda, Junichiro Exp Anim Original We analyzed the Hr gene of a hairless mouse strain of unknown origin (HR strain, http://animal.nibio.go.jp/e_hr.html) to determine whether the strain shares a mutation with other hairless strains, such as HRS/J and Skh:HR-1, both of which have an Hr(hr) allele. Using PCR with multiple pairs of primers designed to amplify multiple overlapping regions covering the entire Hr gene, we found an insertion mutation in intron 6 of mutant Hr genes in HR mice. The DNA sequence flanking the mutation indicated that the mutation in HR mice was the same as that of Hr(hr) in the HRS/J strain. Based on the sequence, we developed a genotyping method using PCR to determine zygosities. Three primers were designed: S776 (GGTCTCGCTGGTCCTTGA), S607 (TCTGGAACCAGAGTGACAGACAGCTA), and R850 (TGGGCCACCATGGCCAGATTTAACACA). The S776 and R850 primers detected the Hr(hr) allele (275-bp amplicon), and S607 and R850 identified the wild-type Hr allele (244-bp amplicon). Applying PCR using these three primers, we confirmed that it is possible to differentiate among homozygous Hr(hr) (longer amplicons only), homozygous wild-type Hr(shorter amplicons only), and heterozygous (both amplicons) in HR and Hos:HR-1 mice. Our genomic analysis indicated that the HR, HRS/J, and Hos:HR-1 strains, and possibly Skh:HR-1 (an ancestor of Hos:HR-1) strain share the same Hr(hr) gene mutation. Our genotyping method will facilitate further research using hairless mice, and especially immature mice, because pups can be genotyped before their phenotype (hair coat loss) appears at about 2 weeks of age. Japanese Association for Laboratory Animal Science 2013-08-01 2013 /pmc/articles/PMC4160947/ /pubmed/23903062 http://dx.doi.org/10.1538/expanim.62.267 Text en ©2013 Japanese Association for Laboratory Animal Science http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. |
spellingShingle | Original Suzuki, Osamu Koura, Minako Noguchi, Yoko Uchio-Yamada, Kozue Matsuda, Junichiro Zygosity Determination in Hairless Mice by PCR Based on Hr(hr) Gene Analysis |
title | Zygosity Determination in Hairless Mice by PCR Based on
Hr(hr) Gene
Analysis |
title_full | Zygosity Determination in Hairless Mice by PCR Based on
Hr(hr) Gene
Analysis |
title_fullStr | Zygosity Determination in Hairless Mice by PCR Based on
Hr(hr) Gene
Analysis |
title_full_unstemmed | Zygosity Determination in Hairless Mice by PCR Based on
Hr(hr) Gene
Analysis |
title_short | Zygosity Determination in Hairless Mice by PCR Based on
Hr(hr) Gene
Analysis |
title_sort | zygosity determination in hairless mice by pcr based on
hr(hr) gene
analysis |
topic | Original |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4160947/ https://www.ncbi.nlm.nih.gov/pubmed/23903062 http://dx.doi.org/10.1538/expanim.62.267 |
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