PMID: 2860679May 1, 1985Paper

Cloning and analysis of the Clostridium perfringens tetracycline resistance plasmid, pCW3

Plasmid
L J Abraham, J I Rood

Abstract

Clostridium perfringens strain CW92 carries pCW3, a conjugative 47-kb plasmid that confers inducible resistance to tetracycline. The plasmid was examined by restriction endonuclease analysis and by cloning each of the five ClaI fragments of pCW3 in Escherichia coli, using pBR322. Analysis of the recombinant plasmids allowed the deduction of a detailed restriction map of pCW3. The tetracycline resistance determinant of pCW3 was mapped by examining the phenotype of recombinant E. coli clones derived from the cloning, into pUC vector plasmids, of EcoRI fragments from pCW3. The C. perfringens tetracycline resistance determinant was expressed in E. coli and was shown to be located on two juxtaposed EcoRI fragments which together encompass a 4-kb region of pCW3. Deletion experiments showed that the tetracycline resistance gene, and/or its control regions, contained internal EcoRI and SphI sites. E. coli strains that carried recombinant plasmids with only the 4-kb region were found to express tetracycline resistance constitutively. In contrast, recombinant plasmids harboring a 10.5-kb ClaI fragment of pCW3, that included the 4-kb region, coded for an inducible tetracycline resistance phenotype. The existence of a negatively regulated ...Continue Reading

References

Nov 1, 1977·Plasmid·J GrinstedM H Richmond
May 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·H L YangS B Levy
Jun 1, 1981·Analytical Biochemistry·D S Holmes, M Quigley
Jan 11, 1984·Nucleic Acids Research·J I Rood, J M Gawthorne

Citations

Dec 1, 1989·FEMS Microbiology Reviews·M YoungW L Staudenbauer
Dec 19, 2003·Avian Pathology : Journal of the W.V.P.A·A MartelF Haesebrouck
Apr 9, 2001·Infection and Immunity·S BrynestadJ I Rood
Nov 22, 2001·Journal of Bacteriology·P A JohanesenJ I Rood
May 24, 2013·Microbiology and Molecular Biology Reviews : MMBR·Jihong LiBruce A McClane

Related Concepts

Genome Mapping
Clostridium perfringens
DNA Restriction Enzymes
Alkalescens-Dispar Group
Genes, Bacterial
R Factors
Topicycline

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