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Identification and Characterization of a New Porin Gene of Klebsiella pneumoniae: Its Role in β-Lactam Antibiotic Resistance

机译:肺炎克雷伯菌新的猪基因的鉴定和表征:其在β-内酰胺抗生素耐药性中的作用

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摘要

Klebsiella pneumoniae porin genes were analyzed to detect mutations accounting for the porin deficiency observed in many β-lactam-resistant strains. PCR and Southern blot analysis revealed the existence of a third porin gene in addition to the OmpK36 and OmpK35 porin genes previously described. This new porin gene was designated ompK37 and is present in all of the clinical isolates tested. The OmpK37 porin gene was cloned, sequenced, and overexpressed in Escherichia coli. In contrast to that of the major porins, OmpK37 porin expression was only detectable by Western blot analysis in porin-deficient β-lactam-resistant strains, suggesting strong down regulation under standard laboratory conditions. Functional characterization suggested a narrower pore for the OmpK37 porin than for K. pneumoniae porins OmpK36 and OmpK35. This correlated with the susceptibility to certain β-lactam antibiotics, since a K. pneumoniae strain expressing porin OmpK37, but not porin OmpK36 or OmpK35, was less susceptible to β-lactam antibiotics than the same strain expressing either porin OmpK36 or OmpK35.
机译:分析了肺炎克雷伯菌的孔蛋白基因,以检测导致在许多对β-内酰胺耐药的菌株中观察到的孔蛋白缺乏的突变。 PCR和Southern印迹分析显示,除了先前描述的OmpK36和OmpK35孔蛋白基因外,还存在第三孔蛋白基因。这个新的孔蛋白基因被命名为ompK37,存在于所有测试的临床分离株中。 OmpK37孔蛋白基因在大肠杆菌中被克隆,测序和过表达。与主要孔蛋白相反,OmpK37孔蛋白表达只能通过蛋白质印迹分析在缺乏孔蛋白的β-内酰胺抗性菌株中检测到,表明在标准实验室条件下强烈下调。功能表征表明,OmpK37孔蛋白的孔比肺炎克雷伯菌孔蛋白OmpK36和OmpK35的孔窄。这与对某些β-内酰胺抗生素的敏感性有关,因为与表达porin OmpK36或OmpK35的相同菌株相比,表达孔蛋白OmpK37而不是孔蛋白OmpK36或OmpK35的肺炎克雷伯菌菌株对β-内酰胺抗生素的敏感性较低。

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