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Abstract
Fragments of plasmid pIP816, which confers high-level glycopeptide resistance in Enterococcus faecium BM4147, were cloned into a conjugative gram-negative-gram-positive shuttle vector. The resulting hybrids were transferred by conjugation from Escherichia coli to Enterococcus faecalis and Bacillus thuringiensis. A 4-kilobase EcoRI fragment from pIP816 was found to confer vancomycin resistance in these hosts but not in E. coli or Bacillus subtilis.
Highlights
Units des Agents Antibactdriens, Unitt Associee Centre National de la Recherche Scientifique 271, Institut Pasteur, 75724 Paris Cedex 15,1 and Service de Bacttriologie-Virologie-Hygihne, HBpital Henri Mondor, Universitt Paris XII, 94010 Crfteil/2 France
We previously established that resistance to high levels of vancomycin and teicoplanin in four clinical isolates of Enterococcus faecium was plasmid mediated and inducible by subinhibitory concentrations of glycopeptide antibiotics [10, 11]
We describe the cloning of the resistance determinant vanA encoding inducible high-level resistance to glycopeptides in E. faecium BM4147 [10]
Summary
Mants resistant to kanamycin were tested for glycopeptide resistance. The plasmid content of the transconjugants and of the transformants was studied by agarose gel electrophoresis of crude bacterial lysates after digestion with EcoRI endonuclease. E.faecalis JH2-2 containing pAT211 expressed glycopeptide resistance at a high level, similar to that of E. faecium BM4147 (Table 1) [10]. Study of the induction of glycopeptide resistance revealed that E. faecalis JH2-2 containing plasmids pAT211 and pAT213 displayed the same pattern of inducibility as did the original E. faecium clinical isolate BM4147 (10; Fig. 3). Whereas vanA confers in E. faecalis the same level of resistance as it does in the original E. faecium host [10], the expression appeared to be lower in B. thuringiensis. To rule out the possibility that lack of expression was due to deletions, plasmids pAT211 and pAT213 were purified from the B. subtilis and E. coli transformants, transformed into E. coli JM83 containing pRK24, and transferred by conjugation into E. faecalis JH2-2.
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