Using a plasmid containing a transcriptional fusion in which the E.coli galK gene is expressed from the trfB promoter of broad host range plasmid RK2 we show that transcription from the trfB promoter is repressed by the products of both the trfB and korB genes as we have previously predicted from the sequence homology of the trfA and trfB promoters and the fact that the trfA promoter is regulated by trfB and korB. These loci, trfB and korB are normally transcribed from the trfB promoter. Thus the trfB incC korB operon of RK2 is doubly autogeneously regulated. In addition, we describe the isolation and characterization of a mutant trfA promoter which has become insensitive to repression by trfB as a result of a point mutation within the inverted repeat sequence previously predicted to be the trfB protein binding site. These results provide strong evidence for our previously proposed model for control of transcription from the trfA and trfB promoters.