Model involving gene inactivation in the generation of autosomal recessive mutants in mammalian cells in culture

A. E. Simon, M. W. Taylor, W. E C Bradley, L. H. Thompson

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

We present evidence for a two-step model for expression of the recessive phenotype at the diploid adenine phosphoribosyl transferase (aprt) locus in Chinese hamster ovary cells. This model proposes a high-frequency event leading to allelic inactivation and a low-frequency event leading to a structural alteration of the APRT protein. Either event can occur first, resulting in two types of heterozygous cells. The proposed model is based on analysis of Chinese hamster ovary presumptive aprt heterozygotes and APRT- mutants, derived by two different laboratories. The major class of heterozygotes (class 1) had approximately 50% parental APRT activity, 50% immunologically precipitable APRT protein, and only wild-type enzyme as based on two-dimensional gel electrophoresis and thermal inactivation studies. We propose that one allele at the aprt locus has been inactivated in these heterozygotes. APRT- mutants derived from any single class 1 heterozygote arose at a low frequency and contained either no immunologically detectable APRT protein or an APRT enzyme which was, in most cases, demonstrably altered. The second class of heterozygotes, consisting of two independent isolates, gave rise to APRT- cells at a high frequency (10-3 to 10-5). These heterozygous cell lines had 50% of parental APRT activity and only wildtype spot, or wild-type and an electrophoretic variant spot, on two-dimensional gels. These aprt heterozygotes appear to have arisen by mutation at one allele. APRT- mutants derived from either heterozygote of this class had all lost the wild-type activity, consistent with the proposed model.

Original languageEnglish (US)
Pages (from-to)1126-1133
Number of pages8
JournalMolecular and Cellular Biology
Volume2
Issue number9
StatePublished - 1982
Externally publishedYes

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Gene Silencing
Heterozygote
Cell Culture Techniques
Adenine
Transferases
Cricetulus
Ovary
Alleles
Proteins
Electrophoresis, Gel, Two-Dimensional
Enzymes
Diploidy
Hot Temperature
Gels
Phenotype
Cell Line
Mutation

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Simon, A. E., Taylor, M. W., Bradley, W. E. C., & Thompson, L. H. (1982). Model involving gene inactivation in the generation of autosomal recessive mutants in mammalian cells in culture. Molecular and Cellular Biology, 2(9), 1126-1133.

Model involving gene inactivation in the generation of autosomal recessive mutants in mammalian cells in culture. / Simon, A. E.; Taylor, M. W.; Bradley, W. E C; Thompson, L. H.

In: Molecular and Cellular Biology, Vol. 2, No. 9, 1982, p. 1126-1133.

Research output: Contribution to journalArticle

Simon, A. E. ; Taylor, M. W. ; Bradley, W. E C ; Thompson, L. H. / Model involving gene inactivation in the generation of autosomal recessive mutants in mammalian cells in culture. In: Molecular and Cellular Biology. 1982 ; Vol. 2, No. 9. pp. 1126-1133.
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