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Molecular Analysis of Mutations in the Human HPRT Gene

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Molecular Toxicology Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2102))

Abstract

The HPRT assay uses incorporation of toxic nucleotide analogues to select for cells lacking the purine scavenger enzyme hypoxanthine-guanine phosphoribosyl transferase. A major advantage of this assay is the ability to isolate mutant cells and determine the molecular basis for their functional deficiency. Many types of analyses have been performed at this locus: the current protocol involves generation of a cDNA and multiplex PCR of each exon, including the intron/exon junctions, followed by direct sequencing of the products. This analysis detects point mutations, small deletions and insertions within the gene, mutations affecting RNA splicing, and the products of illegitimate V(D)J recombination within the gene. Establishment of and comparisons with mutational spectra hold the promise of identifying exposures to mutation-inducing genotoxicants from their distinctive pattern of gene-specific DNA damage at this easily analyzed reporter gene.

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Author information

Authors and Affiliations

  1. Department of Environmental and Occupational Health, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA, USA

    Phouthone Keohavong

  2. Public Health Program, Nova Southeastern University, Fort Lauderdale, FL, USA

    Stephen G. Grant

  3. Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA

    Liqiang Xi

Authors
  1. Phouthone Keohavong
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  2. Liqiang Xi
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  3. Stephen G. Grant
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Corresponding author

Correspondence to Phouthone Keohavong .

Editor information

Editors and Affiliations

  1. Department of Environmental and Occupational Health, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA, USA

    Phouthone Keohavong

  2. Department of Environmental Toxicology, The Institute of Environmental and Human Health (TIEHH), Texas Tech University, Lubbock, TX, USA

    Kamaleshwar P. Singh

  3. Department of Occupational and Environmental Health Sciences, School of Public Health West Virginia University, Morgantown, WV, USA

    Weimin Gao

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Keohavong, P., Xi, L., Grant, S.G. (2020). Molecular Analysis of Mutations in the Human HPRT Gene. In: Keohavong, P., Singh, K., Gao, W. (eds) Molecular Toxicology Protocols. Methods in Molecular Biology, vol 2102. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0223-2_20

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  • DOI: https://doi.org/10.1007/978-1-0716-0223-2_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0222-5

  • Online ISBN: 978-1-0716-0223-2

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