Project Details
Description
The DNA of a cell presents the first line of control over metabolic and
genetic processes. Replication of this repository of information is the
primal biochemical act of cells. The goals of this proposal are to
determine how this event occurs. Our focus will be on the function of DNA polymerases in E. coli. We will
use biochemical and genetic means to analyze a mutation (pcbA) which
permits by-pass of a replication block due to a defective DNA polymerase
III through utilization of DNA polymerase I. We will investigate the
biochemical mechanism of action of the pcbA mutation using in vitro
complementation in a phage replication system. Characterization of the pcb
gene will also require cloning, using complementation of temperature
sensitivity in polCts mutants. Achieving this goal will permit biochemical
analysis of the mutant gene and study of the gene product. The relief of
replication blockade suggests that DNA polymerase III may be eliminated by
mutagenesis. We will attempt to do that. We will also purify and
characterize the DNA polymerase I arising as a reaction of suppression of
the polAI amber mutation.
genetic processes. Replication of this repository of information is the
primal biochemical act of cells. The goals of this proposal are to
determine how this event occurs. Our focus will be on the function of DNA polymerases in E. coli. We will
use biochemical and genetic means to analyze a mutation (pcbA) which
permits by-pass of a replication block due to a defective DNA polymerase
III through utilization of DNA polymerase I. We will investigate the
biochemical mechanism of action of the pcbA mutation using in vitro
complementation in a phage replication system. Characterization of the pcb
gene will also require cloning, using complementation of temperature
sensitivity in polCts mutants. Achieving this goal will permit biochemical
analysis of the mutant gene and study of the gene product. The relief of
replication blockade suggests that DNA polymerase III may be eliminated by
mutagenesis. We will attempt to do that. We will also purify and
characterize the DNA polymerase I arising as a reaction of suppression of
the polAI amber mutation.
| Status | Finished |
|---|---|
| Effective start/end date | 8/1/78 → 2/28/90 |
Funding
- National Institutes of Health
ASJC
- Medicine(all)
- Biochemistry, Genetics and Molecular Biology(all)
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