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Antimicrobial Agents and Chemotherapy, April 2004, p. 1151-1158, Vol. 48, No. 4
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.4.1151-1158.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

CFE-1, a Novel Plasmid-Encoded AmpC ß-Lactamase with an ampR Gene Originating from Citrobacter freundii

Ryuichi Nakano,1 Ryoichi Okamoto,1 Yumiko Nakano,1 Kenichi Kaneko,1 Naohiro Okitsu,1,2 Yoshio Hosaka,1 and Matsuhisa Inoue1*

Department of Microbiology, School of Medicine and Environmental Infectious Disease, Graduate School of Medical Sciences, Kitasato University, Sagamihara, Kanagawa,1 Department of Otolaryngology, Tohoku University School of Medicine, Sendai, Japan2

Received 13 May 2003/ Returned for modification 1 September 2003/ Accepted 30 December 2003

A clinical isolate of Escherichia coli from a patient in Japan, isolate KU6400, was found to produce a plasmid-encoded ß-lactamase that conferred resistance to extended-spectrum cephalosporins and cephamycins. Resistance arising from production of a ß-lactamase could be transferred by either conjugation or transformation with plasmid pKU601 into E. coli ML4947. The substrate and inhibition profiles of this enzyme resembled those of the AmpC ß-lactamase. The resistance gene of pKU601, which was cloned and expressed in E. coli, proved to contain an open reading frame showing 99.8% DNA sequence identity with the ampC gene of Citrobacter freundii GC3. DNA sequence analysis also identified a gene upstream of ampC whose sequence was 99.0% identical to the ampR gene from C. freundii GC3. In addition, a fumarate operon (frdABCD) and an outer membrane lipoprotein (blc) surrounding the ampR-ampC genes in C. freundii were identified, and insertion sequence (IS26) elements were observed on both sides of the sequences identified (forming an IS26 composite transposon); these results confirm the evidence of the translocation of a ß-lactamase-associated gene region from the chromosome to a plasmid. Finally, we describe a novel plasmid-encoded AmpC ß-lactamase, CFE-1, with an ampR gene derived from C. freundii.

* Corresponding author. Mailing address: Department of Microbiology, School of Medicine and Environmental Infectious Disease, Graduate School of Medical Sciences, Kitasato University, Sagamihara 1-15-1, Kanagawa 228-8555, Japan. Phone: 81-42-778-9349. Fax: 81-42-778-9350. E-mail: matsu{at}kitasato-u.ac.jp.

Antimicrobial Agents and Chemotherapy, April 2004, p. 1151-1158, Vol. 48, No. 4
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.4.1151-1158.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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