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Isolation of the Gene Encoding Yeast Single-Stranded Nucleic Acid Binding Protein 1

Jong, Ambrose Y.-S. and Campbell, Judith L. (1986) Isolation of the Gene Encoding Yeast Single-Stranded Nucleic Acid Binding Protein 1. Proceedings of the National Academy of Sciences of the United States of America, 83 (4). pp. 877-881. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:JONpnas86

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Abstract

A yeast gene encoding SSB-1, a single-stranded nucleic acid binding protein, has been isolated by screening a lambda gt11 genomic DNA library. The gene is located on a 1.84-kilobase chromosomal Bgl II-BamHI fragment. Yeast strains carrying the high-copy-number vector YEp24 with an SSB1 gene insert overproduce SSB-1 3-fold and SSB-1 mRNA 10-fold. A typical haploid cell contains about 20,000 molecules of SSB-1; thus, the cells can tolerate up to 60,000 copies. Yeast SSB-1 was expressed in Escherichia coli cells by using a phage T7 expression system. Spores containing the gene disrupted at a point within the coding sequence germinate and grow normally; thus, the gene is not essential. Protein blots show that no SSB-1 or novel immunologically related species that might retain SSB-1 activity are present in cells containing the disrupted SSB1 genes. Southern analysis and protein blots suggest the presence in yeast of a second, related, but nonidentical gene and two immunologically related proteins of 55 kDa and 75 kDa.


Item Type:Article
Additional Information:Copyright © 1986 by the National Academy of Sciences Communicated by John Abelson, September 30, 1985 We thank Eric M. Phizicky for comments on the manuscript and Mary Gilbert for technical assistance. This investigation was supported by Grant GM 25508 from the National Institutes of Health, Grant MV-142 from the American Cancer Society, and Grant 1-794 from the March of Dimes. 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:unwinding protein; gene disruption; λgt11
Record Number:CaltechAUTHORS:JONpnas86
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:JONpnas86
Alternative URL:http://www.pnas.org/cgi/content/abstract/83/4/877
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:5773
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:01 Nov 2006
Last Modified:26 Dec 2012 09:15

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