Isolation of the gene encoding yeast DNA polymerase I

Cell. 1985 Nov;43(1):369-77. doi: 10.1016/0092-8674(85)90042-x.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Cycle
  • Cloning, Molecular
  • Cross Reactions
  • DNA Polymerase I / biosynthesis
  • DNA Polymerase I / genetics*
  • DNA Polymerase I / immunology
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Genes*
  • Genes, Fungal
  • Genetic Vectors
  • Mutation
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics*


  • DNA Polymerase I