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PMID: 2355921 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cloning and analysis of a gene involved in DNA repair and recombination, the rad1 gene of Schizosaccharomyces pombe.

Molecular and cellular biology ·Vol. 10 ·No. 7 ·1990-07-00 ·Pages 3750-60

Sunnerhagen P, Seaton BL, Nasim A, Subramani S

Abstract

We have cloned the rad1 gene of Schizosaccharomyces pombe by complementation of the rad1-1 mutant, which is deficient in DNA repair and recombination. The coding region of the gene is 582 base pairs long and contains no introns. The predicted product is a strongly acidic, 22-kilodalton protein containing 194 amino acid residues. This gene does not exhibit significant homology to any other known repair gene. The major transcription start site is at 27 base pairs upstream of the putative start codon. Insertion mutagenesis revealed that besides the coding region, at least 151 base pairs of 5'-flanking sequence are required for full complementing activity. A strain carrying a null allele of rad1 was constructed and found to have a phenotype closely similar to that of the rad1-1 mutant. Expression in Escherichia coli of the coding region yielded a protein product of a size close to that predicted from the DNA sequence. This product reacted with antibodies raised against a synthetic peptide with a sequence from that predicted for the protein product. We have localized the rad1 gene to NotI fragment E of the S. pombe genome.

MeSH Terms
Amino Acid Sequence Base Sequence Blotting, Southern DNA Repair DNA, Fungal/genetics,isolation & purification Dose-Response Relationship, Radiation Gamma Rays Genes, Viral Genetic Complementation Test Molecular Sequence Data Oligonucleotide Probes RNA, Fungal/genetics,isolation & purification Recombination, Genetic Restriction Mapping Saccharomycetales/genetics Schizosaccharomyces/genetics,radiation effects Ultraviolet Rays
Chemicals
DNA, Fungal Oligonucleotide Probes RNA, Fungal
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Sunnerhagen P
Department of Biology B-022, University of California, San Diego, La Jolla 92093.
Seaton B L
Nasim A
Subramani S
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.7.3750-3760.1990
Published
1990-07-00
Pages
3750-60
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360827
Subset
IM
Grants
NCI NIH HHS · CA01062 · United States
NIGMS NIH HHS · GM31253 · United States
Databases
GENBANK
M38132
External Links
PubMed source DOI: 10.1128/mcb.10.7.3750-3760.1990 PMC Full Text View journal details
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