|Title||Polymerase chain reaction-based comprehensive procedure for the analysis of the mutation spectrum at the hypoxanthine-guanine phosphoribosyltransferase locus in Chinese hamster cells.|
|Publication Type||Journal Article|
|Year of Publication||1992|
|Authors||Yu, YJ, Xu, Z, Gibbs, RA, Hsie, AW|
|Journal||Environ Mol Mutagen|
|Keywords||Animals, Base Sequence, CHO Cells, Cricetinae, DNA, Exons, Hypoxanthine Phosphoribosyltransferase, Molecular Sequence Data, Mutation, Polymerase Chain Reaction|
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 Journal||Environ. Mol. Mutagen.|