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PMID: 8550486 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Development of techniques for the genetic manipulation of the gliding bacterium Cytophaga johnsonae.

Journal of bacteriology ·Vol. 178 ·No. 3 ·1996-02-00 ·Pages 583-90

McBride MJ, Kempf MJ

Abstract

Cytophaga johnsonae displays many features that make it an excellent model of bacterial gliding motility. Unfortunately, genetic analyses of C. johnsonae, or any related gliding bacteria, were not possible because of a complete lack of selectable markers, cloning vectors, transposons, and convenient methods of gene transfer. As a first step toward a molecular analysis of gliding motility of C. johnsonae, we developed these genetic techniques and tools. Common broad-host-range plasmids and transposons did not function in C. johnsonae. We identified one Bacteroides transposon, Tn4351, that could be introduced into C. johnsonae on plasmid R751 by conjugation from Escherichia coli. Tn4351 inserted in the C. johnsonae genome and conferred erythromycin resistance. Tn-4351 insertions resulted in auxotrophic mutations and motility mutations. We constructed novel plasmids and cosmids for genetic analyses of C. johnsonae. These cloning vectors are derived from a small cryptic plasmid (pCP1) that we identified in the fish pathogen Cytophaga psychrophila D12. These plasmids contain the ermF (erythromycin resistance) gene from Tn4351 and a variety of features that facilitate propagation and selection in E. coli and conjugative transfer from E. coli to C. johnsonae.

MeSH Terms
Blotting, Southern Cytophaga/genetics DNA Transposable Elements Electroporation Genetic Vectors Mutagenesis Plasmids
Chemicals
DNA Transposable Elements
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
McBride M J
Department of Biological Sciences, University of Wisconsin--Milwaukee 53201, USA.
Kempf M J
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.178.3.583-590.1996
Published
1996-02-00
Pages
583-90
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC177698
Subset
IM
Grants
NIGMS NIH HHS · GM51010 · United States
External Links
PubMed source DOI: 10.1128/jb.178.3.583-590.1996 PMC Full Text View journal details
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