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Temperature-Sensitive Mutations in the Yeast DNA Polymerase I Gene

Budd, Martin and Campbell, Judith L. (1987) Temperature-Sensitive Mutations in the Yeast DNA Polymerase I Gene. Proceedings of the National Academy of Sciences of the United States of America, 84 (9). pp. 2838-2842. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:BUDpnas87

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Abstract

Seven mutations that yield thermolabile DNA polymerases have been isolated in the DNA polymerase I gene, POL1, of Saccharomyces cerevisiae. Strains carrying the mutant genes are temperature sensitive for growth. The pol1 mutants were identified by a method that has general applicability for identification of both temperature-sensitive and null mutations. A plasmid containing a mutagenized pol1 gene was transformed into a strain in which the only functional copy of the POL1 gene was carried on an unstable plasmid. The genes conferring temperature-sensitive growth were detected after elimination of the unstable plasmid containing the wild-type gene. DNA polymerase I isolated from each of the mutants is defective at both 23 degrees C and 36 degrees C. DNA synthesis is deficient in vivo at 36 degrees C in all the mutants, while RNA synthesis is normal in all but one of the mutants. The terminal phenotype of pol1 temperature-sensitive mutants is dumbbell-shaped cells in which the nucleus has migrated to, but apparently not entered, the isthmus separating the mother and the daughter. The POL1 gene is located on chromosome XIV approx 2 centimorgans away from met4.


Item Type:Article
Additional Information:Copyright © 1987 by the National Academy of Sciences Communicated by John Abelson, December 23, 1986 Support of this research by the National Institutes of Health (GM 25508) is gratefully acknowledged. 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.
Subject Keywords:Saccharomyces cerevisiae; DNA replication; DNA polymerase a; plasmid mutagenesis
Record Number:CaltechAUTHORS:BUDpnas87
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:BUDpnas87
Alternative URL:http://www.pnas.org/cgi/content/abstract/84/9/2838
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:5002
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:18 Sep 2006
Last Modified:26 Dec 2012 09:03

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