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Antimicrobial Agents and Chemotherapy, July 1999, p. 1584-1590, Vol. 43, No. 7
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Cloning and Characterization of blaVIM, a
New Integron-Borne Metallo-
-Lactamase Gene from a Pseudomonas
aeruginosa Clinical Isolate
Laura
Lauretti,1
Maria Letizia
Riccio,1
Annarita
Mazzariol,2
Giuseppe
Cornaglia,2
Gianfranco
Amicosante,3
Roberta
Fontana,2 and
Gian
Maria
Rossolini1,*
Dipartimento di Biologia Molecolare, Sezione
di Microbiologia, Università di Siena,
53100-Siena,1 Istituto di Microbiologia,
Università di Verona, 37134-Verona,2 and
Dipartimento di Scienze e Tecnologie Biomediche e
Biometria, Università dell'Aquila,
67100-L'Aquila,3 Italy
Received 8 October 1998/Returned for modification 30 January
1999/Accepted 17 April 1999
Production of a metallo-
-lactamase activity was detected in a
carbapenem-resistant Pseudomonas aeruginosa clinical
isolate (isolate VR-143/97) from an Italian inpatient at the Verona
University Hospital (northern Italy). The metallo-
-lactamase
determinant was isolated from a genomic library of VR-143/97,
constructed in an Escherichia coli plasmid vector, by
screening for clones with reduced susceptibility to imipenem.
Sequencing of the cloned gene revealed that it encoded a new class B
-lactamase that was named VIM-1. At the sequence level VIM-1 was
rather divergent from the other class B enzymes (16.4 to 38.7%
identity), overall being more similar to members of subclass B1
including the
-lactamase II of Bacillus cereus (Bc-II),
the Bacteroides fragilis CcrA, the Chryseobacterium
meningosepticum BlaB, and the cassette-encoded IMP-1 enzymes.
Among these, VIM-1 showed the highest degree of similarity to Bc-II.
Similarly to blaIMP,
blaVIM was also found to be carried on a gene
cassette inserted into a class 1 integron. The
blaVIM-containing integron was located on the
chromosome of P. aeruginosa VR-143/97, and the
metallo-
-lactamase-encoding determinant was not transferable to
E. coli by conjugation. Expression of the integron-borne
blaVIM gene in E. coli resulted in
a significant decrease in susceptibility to a broad array of
-lactams (ampicillin, carbenicillin, piperacillin, mezlocillin,
cefotaxime, cefoxitin, ceftazidime, cefoperazone, cefepime, and
carbapenems), revealing a very broad substrate specificity of the VIM-1 enzyme.
*
Corresponding author. Mailing address: Dipartimento di
Biologia Molecolare, Sez. di Microbiologia, Università di Siena,
Via Laterina, 8, 53100-Siena, Italy. Phone: 39 0577 233327. Fax: 39 0577 233325. E-mail: rossolini{at}unisi.it.
Antimicrobial Agents and Chemotherapy, July 1999, p. 1584-1590, Vol. 43, No. 7
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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Murphy, T. A., Simm, A. M., Toleman, M. A., Jones, R. N., Walsh, T. R.
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Docquier, J.-D., Lamotte-Brasseur, J., Galleni, M., Amicosante, G., Frere, J.-M., Rossolini, G. M.
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Miriagou, V., Tzelepi, E., Gianneli, D., Tzouvelekis, L. S.
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Pournaras, S., Tsakris, A., Maniati, M., Tzouvelekis, L. S., Maniatis, A. N.
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Mammeri, H., Bellais, S., Nordmann, P.
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Yong, D., Lee, K., Yum, J. H., Shin, H. B., Rossolini, G. M., Chong, Y.
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Yan, J.-J., Ko, W.-C., Chuang, C.-L., Wu, J.-J.
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Bellais, S., Naas, T., Nordmann, P.
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De Champs, C., Poirel, L., Bonnet, R., Sirot, D., Chanal, C., Sirot, J., Nordmann, P.
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Payne, D. J., Hueso-Rodriguez, J. A., Boyd, H., Concha, N. O., Janson, C. A., Gilpin, M., Bateson, J. H., Cheever, C., Niconovich, N. L., Pearson, S., Rittenhouse, S., Tew, D., Diez, E., Perez, P., de la Fuente, J., Rees, M., Rivera-Sagredo, A.
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Dubois, V., Poirel, L., Marie, C., Arpin, C., Nordmann, P., Quentin, C.
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Prats, G., Miro, E., Mirelis, B., Poirel, L., Bellais, S., Nordmann, P.
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Riccio, M. L., Pallecchi, L., Fontana, R., Rossolini, G. M.
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Galleni, M., Lamotte-Brasseur, J., Rossolini, G. M., Spencer, J., Dideberg, O., Frère, J.-M.
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Rossolini, G. M., Condemi, M. A., Pantanella, F., Docquier, J.-D., Amicosante, G., Thaller, M. C.
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Poirel, L., Girlich, D., Naas, T., Nordmann, P.
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Poirel, L., Lambert, T., Türkoglü, S., Ronco, E., Gaillard, J.-L., Nordmann, P.
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Avison, M. B., von Heldreich, C. J., Higgins, C. S., Bennett, P. M., Walsh, T. R.
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