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

Yongjia Yu, Z. Xu, R. A. Gibbs, A. W. Hsie

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

We have established a comprehensive procedure based on the polymerase chain reactian (PCR) to analyze the molecular spectrum of mutatians at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus in Chinese hamster cells. The procedure includes direct sequencing of PCR-amplified hprt cPHA for locating point mutatians 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 far a comprehensive study of the mutation spectrum in a large number of HPRT-deficient Chinese hamster cell mutants.

Original languageEnglish (US)
Pages (from-to)267-273
Number of pages7
JournalEnvironmental and Molecular Mutagenesis
Volume19
Issue number4
StatePublished - 1992
Externally publishedYes

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Keywords

  • direct DNA sequence analysis
  • molecular mutagenesis
  • multiplex PCR
  • splicing errors

ASJC Scopus subject areas

  • Genetics
  • Environmental Science(all)
  • Environmental Chemistry
  • Health, Toxicology and Mutagenesis
  • Genetics(clinical)
  • Toxicology

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