Conjugative transfer can be inhibited by blocking relaxase activity within recipient cells with intrabodies.

摘要: Horizontal transfer of antibiotic resistance genes carried by conjugative plasmids poses a serious health problem. As relaxases are transported to recipient cells during bacterial conjugation, we investigated whether blocking relaxase activity in the cell might inhibit conjugation. For that purpose, used an intrabody approach generating single-chain Fv antibody library against TrwC plasmid R388. Recombinant antibodies were engineered for cytoplasmic expression Escherichia coli and either selected vitro their specific binding TrwC, or vivo ability interfere with conjugation using high-throughput mating assay. Several clones identified showing inhibition R388 upon cell. The epitope recognized one these intrabodies was mapped region containing Tyr-26 involved DNA-processing termination reaction. These findings demonstrate transferred plays important role open new identify inhibitors