Polymerase chain reaction-based comprehensive procedure for the analysis of the mutation spectrum at the hypoxanthine-guanine phosphoribosyltransferase locus in Chinese hamster cells.

TitlePolymerase chain reaction-based comprehensive procedure for the analysis of the mutation spectrum at the hypoxanthine-guanine phosphoribosyltransferase locus in Chinese hamster cells.
Publication TypeJournal Article
Year of Publication1992
AuthorsYu, YJ, Xu, Z, Gibbs, RA, Hsie, AW
JournalEnviron Mol Mutagen
Volume19
Issue4
Pagination267-73
Date Published1992
ISSN0893-6692
KeywordsAnimals, Base Sequence, CHO Cells, Cricetinae, DNA, Exons, Hypoxanthine Phosphoribosyltransferase, Molecular Sequence Data, Mutation, Polymerase Chain Reaction
Abstract

We have established a comprehensive procedure based on the polymerase chain reaction (PCR) to analyze the molecular spectrum of mutations at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus in Chinese hamster cells. The procedure includes direct sequencing of PCR-amplified hprt cDNA for locating point mutations in the expressed coding sequences, multiplex PCR amplification of the hprt exons for screening large deletions, and direct sequencing of PCR-amplified hprt exons and their flanking regions for detecting intronic mutations resulting in mRNA splicing errors. Using this procedure, we have identified different types of mutations among a representative collection of spontaneous and induced HPRT-deficient Chinese hamster cell mutants. This procedure is simple, rapid, accurate, and practical for a comprehensive study of the mutation spectrum in a large number of HPRT-deficient Chinese hamster cell mutants.

Alternate JournalEnviron. Mol. Mutagen.
PubMed ID1600952