Molecular cloning of eucaryotic genes required for excision repair of UV-irradiated DNA: Isolation and partial characterization of the RAD3 gene of Saccharomyces cerevisiae

L. Naumovski, E. C. Friedberg

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The authors describe the molecular cloning of a 6-kilobase (kb) fragment of yeast chromosomal DNA containing the RAD3 gene of S. cerevisiae. When present in the autonomously replicating yeast cloning vector YEp24, this fragment transformed two different UV-sensitive, excision repair-defective rad3 mutants of S. cerevisiae to UV resistance. The same result was obtained with a variety of other plasmids containing a 4.5-kb subclone of the 6-kb fragment. The UV sensitivity of mutants defective in the RAD1, RAD2, RAD4, and RAD14 loci was not affected by transformation with these plasmids. The 4.5-kb fragment was subcloned into the integrating yeast vector YIp5, and the resultant plasmid was used to transform the rad3-1 mutant to UV resistance. Both genetic and physical studies showed that this plasmid integrated by homologous recombination into the rad3 site uniquely. The authors conclude from these studies that the cloned DNA that transforms the rad3-1 mutant to UV resistance contains the yeast chromosomal RAD3 gene. The 4.5-kb fragment was mapped by restriction analysis, and studies on some of the subclones generated from this fragment indicate that the RAD3 gene is at least 1.5 kb in size.

Original languageEnglish (US)
Pages (from-to)323-331
Number of pages9
JournalJournal of Bacteriology
Volume152
Issue number1
StatePublished - 1982

Fingerprint

Molecular Cloning
DNA Repair
Saccharomyces cerevisiae
Plasmids
Yeasts
DNA
Genes
Genetic Vectors
Homologous Recombination

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

@article{18c5fe87772641eeb870fe0a71d4fdb1,
title = "Molecular cloning of eucaryotic genes required for excision repair of UV-irradiated DNA: Isolation and partial characterization of the RAD3 gene of Saccharomyces cerevisiae",
abstract = "The authors describe the molecular cloning of a 6-kilobase (kb) fragment of yeast chromosomal DNA containing the RAD3 gene of S. cerevisiae. When present in the autonomously replicating yeast cloning vector YEp24, this fragment transformed two different UV-sensitive, excision repair-defective rad3 mutants of S. cerevisiae to UV resistance. The same result was obtained with a variety of other plasmids containing a 4.5-kb subclone of the 6-kb fragment. The UV sensitivity of mutants defective in the RAD1, RAD2, RAD4, and RAD14 loci was not affected by transformation with these plasmids. The 4.5-kb fragment was subcloned into the integrating yeast vector YIp5, and the resultant plasmid was used to transform the rad3-1 mutant to UV resistance. Both genetic and physical studies showed that this plasmid integrated by homologous recombination into the rad3 site uniquely. The authors conclude from these studies that the cloned DNA that transforms the rad3-1 mutant to UV resistance contains the yeast chromosomal RAD3 gene. The 4.5-kb fragment was mapped by restriction analysis, and studies on some of the subclones generated from this fragment indicate that the RAD3 gene is at least 1.5 kb in size.",
author = "L. Naumovski and Friedberg, {E. C.}",
year = "1982",
language = "English (US)",
volume = "152",
pages = "323--331",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "1",

}

TY - JOUR

T1 - Molecular cloning of eucaryotic genes required for excision repair of UV-irradiated DNA

T2 - Isolation and partial characterization of the RAD3 gene of Saccharomyces cerevisiae

AU - Naumovski, L.

AU - Friedberg, E. C.

PY - 1982

Y1 - 1982

N2 - The authors describe the molecular cloning of a 6-kilobase (kb) fragment of yeast chromosomal DNA containing the RAD3 gene of S. cerevisiae. When present in the autonomously replicating yeast cloning vector YEp24, this fragment transformed two different UV-sensitive, excision repair-defective rad3 mutants of S. cerevisiae to UV resistance. The same result was obtained with a variety of other plasmids containing a 4.5-kb subclone of the 6-kb fragment. The UV sensitivity of mutants defective in the RAD1, RAD2, RAD4, and RAD14 loci was not affected by transformation with these plasmids. The 4.5-kb fragment was subcloned into the integrating yeast vector YIp5, and the resultant plasmid was used to transform the rad3-1 mutant to UV resistance. Both genetic and physical studies showed that this plasmid integrated by homologous recombination into the rad3 site uniquely. The authors conclude from these studies that the cloned DNA that transforms the rad3-1 mutant to UV resistance contains the yeast chromosomal RAD3 gene. The 4.5-kb fragment was mapped by restriction analysis, and studies on some of the subclones generated from this fragment indicate that the RAD3 gene is at least 1.5 kb in size.

AB - The authors describe the molecular cloning of a 6-kilobase (kb) fragment of yeast chromosomal DNA containing the RAD3 gene of S. cerevisiae. When present in the autonomously replicating yeast cloning vector YEp24, this fragment transformed two different UV-sensitive, excision repair-defective rad3 mutants of S. cerevisiae to UV resistance. The same result was obtained with a variety of other plasmids containing a 4.5-kb subclone of the 6-kb fragment. The UV sensitivity of mutants defective in the RAD1, RAD2, RAD4, and RAD14 loci was not affected by transformation with these plasmids. The 4.5-kb fragment was subcloned into the integrating yeast vector YIp5, and the resultant plasmid was used to transform the rad3-1 mutant to UV resistance. Both genetic and physical studies showed that this plasmid integrated by homologous recombination into the rad3 site uniquely. The authors conclude from these studies that the cloned DNA that transforms the rad3-1 mutant to UV resistance contains the yeast chromosomal RAD3 gene. The 4.5-kb fragment was mapped by restriction analysis, and studies on some of the subclones generated from this fragment indicate that the RAD3 gene is at least 1.5 kb in size.

UR - http://www.scopus.com/inward/record.url?scp=0020456310&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020456310&partnerID=8YFLogxK

M3 - Article

C2 - 6749808

AN - SCOPUS:0020456310

VL - 152

SP - 323

EP - 331

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 1

ER -