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Molecular Toxicology Protocols pp 255–270Cite as

Analysis of In Vivo Mutation in the Hprt and Tk Genes of Mouse Lymphocytes

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1105))

Abstract

Assays measuring mutant frequencies in endogenous reporter genes are used for identifying potentially genotoxic environmental agents and discovering phenotypes prone to genomic instability and diseases, such as cancer. Here, we describe methods for identifying mouse spleen lymphocytes with mutations in the endogenous X-linked hypoxanthine guanine phosphoribosyl transferase (Hprt) gene and the endogenous autosomal thymidine kinase (Tk) gene. The selective clonal expansion of mutant lymphocytes is based upon the phenotypic properties of HPRT- and TK-deficient cells. The same procedure can be utilized for quantifying Hprt mutations in most strains of mice (and, with minor changes, in other mammalian species), while mutations in the Tk gene can be determined only in transgenic mice that are heterozygous for inactivation of this gene. Expanded mutant clones can be further analyzed to classify the types of mutations in the Tk gene (small intragenic mutations vs. large chromosomal mutations) and to determine the nature of intragenic mutation in both the Hprt and Tk genes.

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Authors and Affiliations

  1. Division of Genetic and Reproductive Toxicology, U.S. FDA/NCTR, 3900 NCTR Road, HFT-120, Jefferson, AR, 72079, USA

    Vasily N. Dobrovolsky, Joseph G. Shaddock & Robert H. Heflich

Authors
  1. Vasily N. Dobrovolsky
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  2. Joseph G. Shaddock
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  3. Robert H. Heflich
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Corresponding author

Correspondence to Vasily N. Dobrovolsky .

Editor information

Editors and Affiliations

  1. Occupational Health, University of Pittsburgh Dept. Environmental &, Pittsburgh, Pennsylvania, USA

    Phouthone Keohavong

  2. Dept. of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida, USA

    Stephen G. Grant

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© 2014 Humana Press

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Dobrovolsky, V.N., Shaddock, J.G., Heflich, R.H. (2014). Analysis of In Vivo Mutation in the Hprt and Tk Genes of Mouse Lymphocytes. In: Keohavong, P., Grant, S. (eds) Molecular Toxicology Protocols. Methods in Molecular Biology, vol 1105. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-739-6_20

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  • DOI: https://doi.org/10.1007/978-1-62703-739-6_20

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-738-9

  • Online ISBN: 978-1-62703-739-6

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