Abstract
DNA polymerase III has been recognized as the required replication enzyme in Escherichia coli. The synthesis subunit of DNA polymerase III holoenzyme (α subunit) is encoded by the dnaE gene. We have reported that E. coli cells can survive and grow in the absence of a functional dnaE gene product if DNA polymerase I and the pcbA1 mutation are present. Existing mutations in the dnaE gene have been conditionally defective thermolabile mutations. We report here construction of nonsense mutations in the dnaE gene by use of a temperature-sensitive suppressor mutation to permit survival at the permissive temperature (23°C). Introduction of the pcbA1 mutation eliminated the temperature sensitive phenotype. We confirmed by immunoblotting the lack of detectable α subunit at 43°C.
Original language | English (US) |
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Pages (from-to) | 3139-3143 |
Number of pages | 5 |
Journal | Journal of bacteriology |
Volume | 171 |
Issue number | 6 |
DOIs | |
State | Published - 1989 |
Externally published | Yes |
ASJC Scopus subject areas
- Microbiology
- Molecular Biology