Development of a novel chloramphenicol resistance expression plasmid used for genetic complementation of a fliG deletion mutant in Treponema denticola.
A new expression plasmid containing the fla operon promoter and a staphylococcal chloramphenicol resistance gene, was constructed to help assess the role of fliG in Treponema denticola motility. Deletion of fliG resulted in a nonmotile mutant with a markedly decreased number of flagellar filaments. Wild-type fliG genes from T. denticola and from Treponema pallidum were cloned into this expression plasmid. In both cases, the gene restored the ability of the mutant to gyrate its cell ends and enabled colony spreading in agarose. This shuttle plasmid enables high-level expression of genes in T. denticola and possesses an efficient selectable marker that provides a new tool for treponemal genetics.
David Axelrod Institute for Public Health, Wadsworth Center, New York State, Department of Health, P.O. Box 22002, Albany, NY 12201-2002, USA.
Linda L
LL
Slivienski-Gebhardt
Jacques
J
Izard
William A
WA
Samsonoff
Ronald J
RJ
Limberger
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics*; D002701:Chloramphenicol / Q000494:pharmacology; D024881:Drug Resistance, Bacterial; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D017422:Sequence Analysis, DNA; D014208:Treponema / Q000187:drug effects / Q000235:genetics / Q000502:physiology*
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; C075660:Flig protein, Bacteria; D002701:Chloramphenicol