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Purification of the cdc8 protein of Saccharomyces cerevisiae by complementation in an aphidicolin-sensitive in vitro DNA replication system

Kuo, Chia-Lam and Campbell, Judith L. (1982) Purification of the cdc8 protein of Saccharomyces cerevisiae by complementation in an aphidicolin-sensitive in vitro DNA replication system. Proceedings of the National Academy of Sciences of the United States of America, 79 (14). pp. 4243-4247. ISSN 0027-8424. PMCID PMC346646. https://resolver.caltech.edu/CaltechAUTHORS:KUOpnas82

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Abstract

DNA synthesis in vitro in Brij-treated Saccharomyces cerevisiae requires the product of the CDC8 gene (Hereford, L. M. & Hartwell, L. H. (1971) Nature (London) New Biol. 234, 171-172). Extracts of wild-type A364a yeast restore DNA synthesis in Brij-treated cdc8, a mutant containing a thermolabile cdc8 gene product. This constitutes a complementation assay by which the cdc8 gene product can be monitored during purification. A heat-stable protein responsible for this complementation has been partially purified from both wild-type A364a cells and from a cdc8 temperature-sensitive mutant. The complementation activity from the mutant is thermolabile when compared to the wild-type activity, indicating that CDC8 is the structural gene for the protein.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC346646/PubMed CentralArticle
Additional Information:© 1982 by the National Academy of Sciences. Communicated by Norman Davidson, July 27, 1981. We thank S.E. Celniker for helpful discussion. This investigation was supported by U.S. Public Health Service Grant GM 25508 and Research Career Development Award CA 00544. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Funders:
Funding AgencyGrant Number
NIHGM 25508
NIHCA 00544
Subject Keywords:permeabilizeds cells; yeast; cell division cycle (cdc) mutants; DNA elongation; eukaryotic DNA replication
Issue or Number:14
PubMed Central ID:PMC346646
Record Number:CaltechAUTHORS:KUOpnas82
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:KUOpnas82
Alternative URL:http://www.pnas.org/cgi/content/abstract/79/14/4243
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:8572
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:21 Aug 2007
Last Modified:02 Oct 2019 23:52

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