Molecular Analysis of Mutations in the Human HPRT Gene

  • Phouthone KeohavongEmail author
  • Liqiang Xi
  • Stephen G. Grant
Part of the Methods in Molecular Biology book series (MIMB, volume 2102)


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.

Key words

HPRT Mutation Mutational spectra Deletions Recombination Mutational fingerprint 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Phouthone Keohavong
    • 1
    Email author
  • Liqiang Xi
    • 3
  • Stephen G. Grant
    • 2
  1. 1.Department of Environmental and Occupational Health, Graduate School of Public HealthUniversity of PittsburghPittsburghUSA
  2. 2.Public Health ProgramNova Southeastern UniversityFort LauderdaleUSA
  3. 3.Center for Cancer ResearchNational Cancer InstituteBethesdaUSA

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