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Antimicrobial Agents and Chemotherapy, September 2004, p. 3284-3290, Vol. 48, No. 9
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.9.3284-3290.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Functional and Structural Characterization of the Genetic Environment of an Extended-Spectrum ß-Lactamase blaVEB Gene from a Pseudomonas aeruginosa Isolate Obtained in India

Daniel Aubert,1 Delphine Girlich,1 Thierry Naas,1 Shanta Nagarajan,2 and Patrice Nordmann1*

Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, Paris, France,1 Department for Microbiology, Batra Hospital and Medical Research Centre, New Delhi, India2

Received 23 January 2004/ Returned for modification 28 March 2004/ Accepted 20 May 2004

A Pseudomonas aeruginosa clinical strain isolated from a patient hospitalized in a New Delhi, India, hospital was resistant to expanded-spectrum cephalosporins, imipenem, and aztreonam. A blaVEB-1-like gene named blaVEB-1a, which codes for the extended-spectrum ß-lactamase VEB-1a, was identified. The genetic environment of blaVEB-1a was peculiar: (i) no 5' conserved sequence (5'-CS) region was present upstream of the ß-lactamase gene, whereas blaVEB-1-like genes are usually associated with class 1 integrons; (ii) blaVEB-1a was inserted between two truncated 3'-CS regions in a direct repeat; and (iii) four 135-bp repeated DNA sequences (repeated elements) were located on each side of the blaVEB-1a gene. Expression of the blaVEB-1a gene was driven by a strong promoter located in one of these repeated sequences. In addition, cloning of the ß-lactamase content of this P. aeruginosa isolate followed by expression in Escherichia coli identified the naturally occurring AmpC ß-lactamase and a gene encoding an OXA-2-like ß-lactamase located in a class 1 integron, In78, in which an insertion sequence, ISpa7, was inserted within its 5'-CS region.

* Corresponding author. Mailing address: Service de Bactériologie-Virologie, Hôpital de Bicêtre, 78 rue du Général Leclerc, 94275 Le Kremlin-Bicêtre Cedex, France. Phone: 33-1-45-21-36-32. Fax: 33-1-45-21-63-40. E-mail: nordmann.patrice{at}bct.ap-hop-paris.fr.

Antimicrobial Agents and Chemotherapy, September 2004, p. 3284-3290, Vol. 48, No. 9
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.9.3284-3290.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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