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《Journal of Jilin Agricultural University》 2003-01
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Cloning and Sequence Mutation Analysis of Quinolone Resistance Determining Regions of Pathogenic Aeromonas hydrophil

LU Qiang1, ZHENG Wei1, LI Lianrui2, LIU Mingyuan1, YU Haibin3(1.Faculty of Animal Science and Veterinary Medicine, The Quartermaster University of PLA, Changchun,Jilin 130062,China; 2.Institute of Animal Science and Technology, Tarim University of Agriculture Reclamation, Alar,Xinjiang 843300; 3.Animal Checking Department of Jiutai,Jiutai,Jilin 130500,China)  
In order to invesgate the mechanism of quinolones drug resistance of Aeromonas hydrophil, the genome of four drug resistant strains, two susceptible strains isolated in Jilin Province were amplified by PCR. The primers, used included A1:AGAGTTCCTATCTTGATTACG;A2:CTGTGATGTAGGTCATCAACT, which were synthesized according to the gyr A gene sequence of A. salmonicida, which amplify the quinolones resistancedetermining region(QRDR) of the gyrA gene. PCR products were cloned into vector pMD18T, the recombinant vectors were transfered into the E.coli DH5αcompetent cells. After identification, the sequences of the amplified QRDRs were determined and analysised by DNA star software. Four mutation sites of amino acid sequence were found, which were serine 83toisoleucine, leucine 92tomethionine, isoleucine 174tophenylalanine and an adjacent site asparagines 202, leucine 203toasparatic acid, arginine. Among 4 mutation sites, only the mutation of site 83 was identical with E.coli and other drug resistant bacteria, the molecular weigh of the mutated amino acid residue was bigger than original residue. The QRDR sequences of four drug resistant strains and two susceptible strains were registered to Genebank with accession numbers being AY039655, AY039656, AY138539,AY138540,AY138537,AY138538.
【Fund】: 国家自然科学基金资助项目(30070587);; 吉林省自然科学基金资助项目(20020648)
【CateGory Index】: S917.1
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