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Mol Genet Genomics ; 270(1): 94-102, 2003 Oct.
Article in English | MEDLINE | ID: mdl-12942368

ABSTRACT

Conjugative transfer of F-like plasmids is a tightly regulated process. The TraJ protein is the main positive activator of the tra operon which encodes products required for conjugative transfer of F-like plasmids. Nucleotide sequence analysis revealed potential Lrp and H-NS binding sites in the traJ regulatory region. Expression of a traJ-lacZ fusion in hns and lrp mutant strains showed that both are positive modulators of traJ expression. Competitive RT-PCR demonstrated that H-NS and Lrp exert their effect at the transcriptional level. Electrophoretic mobility-shift assays showed that H-NS and Lrp proteins bind to the traJ promoter. Conjugative transfer of pRK100 was decreased in hns but not in lrp mutant strains. Together, the results indicate H-NS and Lrp function as activators of traJ transcription.


Subject(s)
Bacterial Proteins/genetics , DNA-Binding Proteins/genetics , Escherichia coli Proteins/genetics , Escherichia coli/genetics , Plasmids/genetics , Proteins/genetics , Transcription Factors , Base Sequence , Crosses, Genetic , Gene Expression Regulation, Bacterial/genetics , Kinetics , Leucine-Responsive Regulatory Protein , Molecular Sequence Data , Proteins/metabolism , RNA, Bacterial/genetics , RNA, Bacterial/isolation & purification , Recombinant Fusion Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , beta-Galactosidase/genetics
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