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Isolation of the gene encoding yeast DNA polymerase I.

Authors
  • Johnson, L M
  • Snyder, M
  • Chang, L M
  • Davis, R W
  • Campbell, J L
Type
Published Article
Journal
Cell
Publisher
Elsevier
Publication Date
Nov 01, 1985
Volume
43
Issue
1
Pages
369–377
Identifiers
PMID: 3907855
Source
Medline
License
Unknown

Abstract

A yeast genomic DNA expression library in lambda gt11 antibody prepared against yeast DNA polymerase I were used to isolate the gene encoding DNA polymerase I. The identity of the DNA polymerase I gene was determined by several criteria. First, the clone-encoded protein is immunologically related to DNA polymerase I. Second, cells containing the gene cloned in the high copy number plasmid YEp24 overproduce the polymerase activity 4- to 5-fold as measured in yeast extracts. Finally, insertion of the gene downstream from a bacteriophage T7 promoter allows synthesis of yeast DNA polymerase I in Escherichia coli. Gene disruption and Southern hybridization experiments show that the polymerase is encoded by an essential, single copy gene. Examination of the germinated spores containing the disrupted gene reveals a defect in nuclear division and a terminal phenotype typical of replication mutants.

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