Molecular characterization of a new class 3 integron in Klebsiella pneumoniae.
Klebsiella pneumoniae FFUL 22K was isolated in April 1999 from the urine of an intensive care unit patient in Portugal. The strain showed an extended-spectrum cephalosporin resistance profile. A typical synergistic effect between cefotaxime or cefepime and clavulanic acid was observed. An Escherichia coli transformant displayed a similar resistance phenotype and harbored a ca. 9.4-kb plasmid (p22K9). Cloning experiments revealed that the extended-spectrum beta-lactamase was encoded by bla(GES-1), previously described in class 1 integrons from K. pneumoniae ORI-1 and Pseudomonas aeruginosa Pa695. Further sequence analysis demonstrated that the bla(GES-1) gene cassette was located on a new class 3 integron. The integron was 2863 bp long and consisted of an intI3 integrase gene, an attI3 recombination site, two promoter regions, and two gene cassettes. The IntI3 integrase was 98.8% identical to that of Serratia marcescens AK9373. The bla(GES-1) gene cassette was inserted at the attI3 site. The second gene cassette was the result of a fusion event between bla(OXA-10)-type and aac(6')-Ib gene cassettes and conferred resistance to kanamycin. This is the second class 3 integron reported and the first time that the bla(GES-1) gene cassette has been found on an integron belonging to this class, highlighting the considerable heterogeneity of their genetic environment and the spread of gene cassettes among different classes of integrons.
Correia
Mário
M
Laboratory of Microbiology, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal.
Boavida
Filipa
F
Grosso
Filipa
F
Salgado
M J
MJ
Lito
L M
LM
Cristino
J Melo
JM
Mendo
Sónia
S
Duarte
Aida
A
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001483:Base Sequence; D018550:Cephalosporin Resistance; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D032023:Integrons / Q000235:genetics*; D007525:Isoelectric Focusing; D007710:Klebsiella Infections / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014169:Transformation, Bacterial; D001618:beta-Lactamases / Q000235:genetics
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D001618:beta-Lactamases