Rapid method for isolation and screening of cytochrome c oxidase-deficient mutants of Saccharomyces cerevisiae
10.1128/jb.161.3.831-835.1985
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

We describe here a new method for the specific isolation of cytochrome c oxidase-deficient mutants of Saccharomyces cerevisiae. One unique feature of the method is the use of tetramethyl-p-phenylenediamine as a cytochrome c oxidase activity stain for yeast colonies. The staining of yeast colonies by tetramethyl-p-phenylenediamine is dependent upon a functional cytochrome c oxidase and is unaffected by other lesions in respiration. Since the tetramethyl-p-phenylenediamine colony staining reaction is rapid and simple, it greatly facilitates both the identification and characterization of cytochrome c oxidase-deficient mutants. Another feature of the method, which is made possible by the tetramethyl-p-phenylenediamine colony stain, is the use of an op1 parent strain for the isolation of nuclear pet or mitochondrial mit mutants in specific protein-coding genes. A parent strain that carries this marker selects against rho0 or rho- classes of pleiotropic respiratory-deficient mutants, since these are lethal in op1 strains. We have used this method to isolate 123 independently derived cytochrome c oxidase-deficient pet mutants and 300 independently derived mit mutants.

Bibliography

McEwen, J. E., Cameron, V. L., & Poyton, R. O. (1985). Rapid method for isolation and screening of cytochrome c oxidase-deficient mutants of Saccharomyces cerevisiae. Journal of Bacteriology, 161(3), 831–835.

Authors 3
  1. J E McEwen (first)
  2. V L Cameron (additional)
  3. R O Poyton (additional)
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Dates
Type When
Created 5 years, 7 months ago (Jan. 3, 2020, 11:54 a.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:42 p.m.)
Indexed 2 months, 3 weeks ago (June 3, 2025, 12:58 a.m.)
Issued 40 years, 5 months ago (March 1, 1985)
Published 40 years, 5 months ago (March 1, 1985)
Published Print 40 years, 5 months ago (March 1, 1985)
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@article{McEwen_1985, title={Rapid method for isolation and screening of cytochrome c oxidase-deficient mutants of Saccharomyces cerevisiae}, volume={161}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.161.3.831-835.1985}, DOI={10.1128/jb.161.3.831-835.1985}, number={3}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={McEwen, J E and Cameron, V L and Poyton, R O}, year={1985}, month=mar, pages={831–835} }