Somatic Cell Genetics

, Volume 3, Issue 6, pp 639–647 | Cite as

Isolation of a lactic dehydrogenase-A-deficient CHO-K1 mutant by nylon cloth replica plating

  • T. D. Stamato
  • Carol Jones
Article
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Revised:

Abstract

A mutant Chinese hamster ovary cell deficient in lactate dehydrogenase A activity has been isolated using a nonselective technique. The method uses histochemical staining to examine colonies directly for enzyme activity and nylon cloth replica plating to recover particular clones. The mutant cell has an apparent Km (pyruvate to lactate) that is nearly tenfold higher than the parental cell, while itsVmax has been reduced more than 80-fold. In mutant cell extracts with added porcine LDH-B enzyme, molecular dissociation and recombination of subunits produces two new active LDH tetramers (A1B3, A2B2). The electrophoretic mobility of at least one of the tetramers (A1B3) was different from those formed in the parental extracts. The evidence suggests the variant cell contains a mutation in the structural gene for LDH-A.

Keywords

Lactate Pyruvate Lactate Dehydrogenase Cell Extract Electrophoretic Mobility 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Plenum Publishing Corporation 1977

Authors and Affiliations

  • T. D. Stamato
    • 1
  • Carol Jones
    • 1
  1. 1.Eleanor Roosevelt Institute for Cancer Research, Department of Biophysics and GeneticsUniversity of Colorado Medical CenterDenver

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