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Cloning and analysis of the Clostridium perfringens tetracycline resistance plasmid, pCW3
Journal article   Peer reviewed

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

L.J. Abraham and J.I. Rood
Plasmid, Vol.13(3), pp.155-162
1985
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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 resistance gene, similar to that proposed for several other bacteria is postulated.

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Citation topics
1 Clinical & Life Sciences
1.120 Inflammatory Bowel Diseases & Infections
1.120.1133 Clostridium Infections
Web Of Science research areas
Genetics & Heredity
Microbiology
ESI research areas
Molecular Biology & Genetics
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