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Characterization of the aac(6')-Ik gene of Acinetobacter sp. 6
Authors:Eric Rudant  Pierre Bourlioux  Patrice Courvalin  Thierry Lambert
Institution:Laboratoire de Bactériologie, Facultéde Médecine Pitié-Salpêtrière, UniversitéPierre et Marie Curie, 91 boulevard de l'hôpital, 75634 Paris Cedex 13, France
Abstract:Abstract Chromosomal DNA of different species of mycobacteria, Mycobacterium tuberculosis, Mycobacterium leprae, Mycobacterium avium and Mycobacterium smegmatis , has been submitted to polymerase chain reaction using two oligonucleotide primers highly homologous to DNA sequences flanking the quinolone resistance-determining region in the gyrA gene of Escherichia coli and Staphylococcus aureus . For each of these mycobacterial species, a 150-bp DNA fragment hybridizing with an intragenic probe of the gyrA gene of E. coli K12 was obtained. The nucleotide sequences of the 108-bp fragments amplified from M. tuberculosis and M. avium were determined. The two sequences were 87% homologous. Except for one residue, their deduced amino acid sequences were identical and shared 67% homology with the quinolone resistance-determining region of the gyrase A subunits of E. coli and S. aureus . Sequencing of the 108-bp fragment amplified from an in vitro mutant of M. avium , highly resistant to fluoroquinolones, showed a point mutation leading to the substitution of Ala for Val at a position corresponding to residues involved in quinolone resistance in E. coli and S. aureus , i.e. Ser 83 for E. coli and Ser 84 for S. aureus .
Keywords:gyrA Gene  DNA gyrase              Mycobacterium            Quinolone  Polymerase chain reaction
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