RESUMEN

Using bioinformatics approaches, 34 potential multidrug resistance (MDR) transporter sequences representing 4 different transporter families were identified in the unannotated Enterococcus faecalis database (TIGR). A functional genomics campaign generating single-gene insertional disruptions revealed several genes whose absence confers significant hypersensitivities to known antimicrobials. We constructed specific strains, disrupted in a variety of previously unpublished, putative MDR transporter genes, as tools to improve the success of whole-cell antimicrobial screening and discovery. Each of the potential transporters was inactivated at the gene level and then phenotypically characterized, both with single disruption mutants and with 2-gene mutants built upon a delta norA deleted strain background.

Asunto(s)

Antibacterianos/síntesis química , Antibacterianos/farmacología , Proteínas Portadoras/metabolismo , Farmacorresistencia Microbiana , Enterococcus faecalis/fisiología , Transportadoras de Casetes de Unión a ATP/genética , Transportadoras de Casetes de Unión a ATP/metabolismo , Antibacterianos/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas Portadoras/genética , Bases de Datos como Asunto , Diseño de Fármacos , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecalis/genética , Eliminación de Gen , Genómica , Proteínas Asociadas a Resistencia a Múltiples Medicamentos , Mutagénesis , Relación Estructura-Actividad