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Antimicrobial Agents and Chemotherapy, January 2003, p. 19-26, Vol. 47, No. 1
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.1.19-26.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Integration of a Transposon Tn1-Encoded Inhibitor-Resistant ß-Lactamase Gene, blaTEM-67 from Proteus mirabilis, into the Escherichia coli Chromosome

Thierry Naas,1* Marie Zerbib,2 Delphine Girlich,1 and Patrice Nordmann1

Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique-Hôpitaux de Paris, Faculté de Médecine Paris-Sud, 94275 Le Kremlin-Bicêtre,1 Unité d'Hygiène, Hôpital Foch, 92151 Suresnes, France2

Received 20 February 2002/ Returned for modification 4 June 2002/ Accepted 21 September 2002

Proteus mirabilis NEL-1 was isolated from a urine sample of a patient hospitalized in a long-term care facility. Strain NEL-1 produced a ß-lactamase with a pI of 5.2 conferring resistance to amoxicillin and amoxicillin-clavulanic acid. Sequencing of a PCR amplicon by using TEM-specific primers revealed a novel blaTEM gene, blaTEM-67. TEM-67 was an IRT-1-like TEM derivative related to TEM-65 (Lys39, Cys244) with an additional Leu21Ile amino acid substitution in the leader peptide. The biochemical properties of TEM-67 were equivalent to those described for TEM-65. Analysis of sequences surrounding blaTEM-67 revealed that it was located on a transposon, Tn1, which itself was located on a 48-kb non-self-transferable plasmid, pANG-1. Electroporation of plasmid pANG-1 into Escherichia coli DH10B resulted in the integration of blaTEM-67 into the chromosome, whereas it remained episomal in the P. mirabilis CIP103181 reference strain. Further characterization of pANG-1 revealed the presence of two identical sequences on both sides of Tn1 that contained an IS26 insertion sequence followed by a novel colicin gene, colZ, which had 20% amino acid identity with other colicin genes. The characterization of this novel TEM derivative provides further evidence for the large diversity of plasmid-encoded ß-lactamases produced in P. mirabilis and for their spread to other enterobacterial species through transposable-element-mediated events.

* Corresponding author. Mailing address: Service de Bactériologie-Virologie, Hôpital de Bicêtre, 78 rue du Général Leclerc 94275 Le Kremlin-Bicêtre Cédex, France. Phone: 33-1-45-21-36-24. Fax: 33-1-45-21-63-40. E-mail: thierry.naas{at}bct.ap-hop-paris.fr.

Antimicrobial Agents and Chemotherapy, January 2003, p. 19-26, Vol. 47, No. 1
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.1.19-26.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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