We will continue our studies on control of plasmid DNA replication in E. coli using P1, a bacteriophage whose prophage is a plasmid, as a model system. From P1 carrying multiple antibiotic resistance determinants, we have isolated mutants that are maintained at multiple copies per cell. We plan to analyze these in detail to determine the molecular mechanisms involved in copy number regulation. We will also isolate mutants that overproduce a factor we believe to be required for P1 plasmid replication and characterize the product they define. We will study the interaction of the different regulatory products by suppressor analysis. We plan to identify and characterize the RNAs and proteins involved in the regulation of replication of this model plasmid and to begin work on an in vitro replication system. In addition, we will complete our work on the generation of a free resistance determinant from P1ApCm.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI017696-08
Application #
3127394
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1981-04-01
Project End
1990-03-31
Budget Start
1988-04-01
Budget End
1990-03-31
Support Year
8
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Emory University
Department
Type
Schools of Medicine
DUNS #
042250712
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Froehlich, B J; Scott, J R (1990) Second-site revertants of the P1 copN22 copy mutant. J Bacteriol 172:2762-4
Froehlich, B J; Scott, J R (1988) A single amino acid difference between Rep proteins of P1 and P7 affects plasmid copy number. Plasmid 19:121-33
Froehlich, B J; Watkins, C; Scott, J R (1986) IS1-dependent generation of high-copy-number replicons from bacteriophage P1 Ap Cm as a mechanism of gene amplification. J Bacteriol 166:609-17