Complementation of a Treponema denticola flgE mutant with a novel coumermycin A1-resistant T. denticola shuttle vector system.
By using the mutated gyrB gene from a spontaneous coumermycin A1-resistant Treponema denticola, an Escherichia coli-T. denticola shuttle vector that renders T. denticola resistant to coumermycin was constructed. The complete T. denticola flgE gene was cloned into the shuttle vector pKMCou, and the vector was transformed into the T. denticola ATCC 33520 flgE erythromycin-resistant knockout mutant HL210. The coumermycin-resistant transformants were motile and restored FlgE activity. This complementation system should prove useful in studying the virulence factors of T. denticola and uncultivatible pathogenic spirochetes.
Chi
Bo
B
Department of Oral Biology, State University of New York at Buffalo, Buffalo, New York 14214, USA.
Limberger
Ronald J
RJ
Kuramitsu
Howard K
HK
D049933:Aminocoumarins; D001426:Bacterial Proteins / Q000235:genetics*; D003374:Coumarins / Q000494:pharmacology*; D027081:DNA Gyrase / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D009154:Mutation; D010957:Plasmids; D059005:Topoisomerase II Inhibitors; D014169:Transformation, Bacterial; D014208:Treponema / Q000187:drug effects / Q000235:genetics* / Q000472:pathogenicity; D014774:Virulence
D049933:Aminocoumarins; D001426:Bacterial Proteins; D003374:Coumarins; D004791:Enzyme Inhibitors; C097681:FlgE protein, Bacteria; D059005:Topoisomerase II Inhibitors; D027081:DNA Gyrase; C004628:coumermycin