An F-derived conjugative cosmid: analysis of tra polypeptides in cosmid-infected cells

The genes involved in the conjugational transfer of F plasmid DNA are organized into three closely linked operons spanning an overall length of approximately 33 kilobase pairs of F. The entire transfer (tra) region comprising all three operons has been cloned into the cosmid vector pHC79 by in vitro recombination and packaging techniques. The transfer-proficient chimeric cosmid pRS2405 was packaged into lambda capsids, and uv-irradiated E. coli cells were infected with these DNA-filled particles. A number of polypeptides programmed by the infecting DNA were identified as tra-specified products; a traJ90 mutation on pRS2405 resulted in the significant reduction of synthesis of all detectable pRS2405-specified tra polypeptides, with the exception of TraTp.