Development of a Novel Chloramphenicol Resistance Expression Plasmid Used for Genetic Complementation of a fliG Deletion Mutant in Treponema denticola

Authors

Linda L. Slivienski-Gebhardt, David Axelrod Institute for Public Health
Jacques Izard, David Axelrod Institute for Public HealthFollow
William A. Samsonoff, David Axelrod Institute for Public Health
Ronald J. Limberger, David Axelrod Institute for Public HealthFollow

Date of this Version

6-9-2004

Citation

2004, American Society for Microbiology. All Rights Reserved

Comments

INFECTION AND IMMUNITY, Sept. 2004, p. 5493–5497 Vol. 72, No. 9 0019-9567/04/$08.000 DOI: 10.1128/IAI.72.9.5493–5497.2004

Abstract

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.