脆弱拟杆菌编码DNA促旋酶的gyrA和gyrB基因的分子克隆。
Molecular cloning of the gyrA and gyrB genes of Bacteroides fragilis encoding DNA gyrase.
作者信息
Onodera Y, Sato K
机构信息
New Product Research Laboratories I, Daiichi Pharmaceutical Co., Ltd., Edogawa-ku, Tokyo 134-8630, Japan.
出版信息
Antimicrob Agents Chemother. 1999 Oct;43(10):2423-9. doi: 10.1128/AAC.43.10.2423.
Abstract
The genes encoding the DNA gyrase A and B subunits of Bacteroides fragilis were cloned and sequenced. The gyrA and gyrB genes code for proteins of 845 and 653 amino acids, respectively. These proteins were expressed in Escherichia coli, and the combination of GyrA and GyrB exhibited ATP-dependent supercoiling activity. To analyze the role of DNA gyrase in quinolone resistance of B. fragilis, we isolated mutant strains by stepwise selection for resistance to increasing concentrations of levofloxacin. We analyzed the resistant mutants and showed that Ser-82 of GyrA, equivalent to resistance hot spot Ser-83 of GyrA in E. coli, was in each case replaced with Phe. These results suggest that DNA gyrase is an important target for quinolones in B. fragilis.
摘要
对脆弱拟杆菌的DNA促旋酶A和B亚基的编码基因进行了克隆和测序。gyrA和gyrB基因分别编码含845和653个氨基酸的蛋白质。这些蛋白质在大肠杆菌中表达，GyrA和GyrB的组合表现出ATP依赖性超螺旋活性。为了分析DNA促旋酶在脆弱拟杆菌对喹诺酮类耐药性中的作用，我们通过逐步选择对浓度递增的左氧氟沙星耐药的菌株来分离突变株。我们对耐药突变株进行了分析，结果表明，GyrA的第82位丝氨酸（相当于大肠杆菌中GyrA的耐药热点第83位丝氨酸）在每种情况下都被苯丙氨酸取代。这些结果表明，DNA促旋酶是喹诺酮类药物作用于脆弱拟杆菌的一个重要靶点。
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