Journal of Antimicrobial Chemotherapy (2000) 45, 783-788
© 2000 The British Society for Antimicrobial Chemotherapy
Analysis of the effects of –42 and –32 ampC promoter mutations in clinical isolates of Escherichia coli hyperproducing AmpC
a Laboratoire de Bactériologie–Virologie, Faculté de Pharmacie, 1 rue Gaston Veil, 44035 Nantes; b Laboratoire de Bactériologie, Virologie, Hygiène hospitalière, CHU, Nantes, France
Escherichia coli usually produces only very small amounts of a constitutive AmpC ß-lactamase, but clinical strains overproducing this enzyme have been isolated. Three different ampC promoters of E. coli clinical strains were cloned upstream of the chloramphenicol acetyltransferase (CAT) gene in the pKK232-8 reporter plasmid and their relative strengths were compared by two different methods. The strength of the promoters from AmpC hyperproducers was 70- to 120-fold higher than those from a low-level AmpC producer. One of the strong promoters, which differs from strain K12 at bases –88, –82, –42, –18, –1 and +58, was mutated to abolish the –42 mutation. This change resulted in a 43-fold decrease in CAT concentration. In another promoter, with eight different mutations at positions –88, –82, –32, –18, –1, +5, +24 and +58, the –32T
A transversion, which created perfect homology with the –35 consensus sequence, was reverted; this led to a 13-fold decrease in CAT concentration. The –42 and –32 mutations play an important role in E. coli resistance to ß-lactams by increasing ampC transcription.
* Corresponding author. Tel: +33-2-40412952; Fax: +33-2-40083829; E-mail: interepi{at}chu-nantes.fr
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