Kim Hong, Kim Sun-Hyun, Ying Yuan-Hai, Kim Hyun-Ju, Koh Young-Hwan, Kim Chang-Jin, Lee Sueng-Hyun, Cha Chang-Yong, Kook Yoon-Hoh, Kim Bum-Joon
Department of Microbiology, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Republic of Korea.
Syst Appl Microbiol. 2005 Jul;28(5):398-404. doi: 10.1016/j.syapm.2005.02.009.
In a previous phylogenetic study of the genus Streptomyces using the rpoB gene, N531, which stands for an aspargine residue in position 531 of RpoB instead of serine (S531), known to be associated with natural rifampin resistance in several organisms, was also observed in the RpoB of several Streptomyces species. To determine whether N531 is associated with the rifampin resistance of Streptomyces strains, we analyzed the rifampin minimum inhibitory concentrations (MICs) of 11 strains of the N531 RpoB type (putative rifampin resistant strains) and of 12 strains of the S531 RpoB type. (putative rifampin susceptible strains). In general, the N531 RpoB types showed higher MIC levels (16-128 microg/ml) than the S531 RpoB types (0-8 microg/ml). To determine the isolation frequencies of N531 RpoB types versus rifampin concentration, we applied screening methods involving different rifampin concentrations (0, 20 and 100 microg/ml) to Korean soils. Higher isolation frequencies of the N531 RpoB types were observed at the higher rifampin concentrations. In addition, during the course of this study we developed an allele specific PCR method to detect rifampin resistant Streptomyces strains. Our results strongly suggested that N531 might be involved in a major mechanism of natural rifampin resistance in strains of the genus Streptomyces.
在先前一项利用rpoB基因对链霉菌属进行的系统发育研究中,在几种链霉菌物种的RpoB中也观察到了N531,它代表RpoB第531位的天冬酰胺残基,而非丝氨酸(S531),已知在几种生物体中与天然利福平抗性相关。为了确定N531是否与链霉菌菌株的利福平抗性有关,我们分析了11株N531 RpoB类型(推定的利福平抗性菌株)和12株S531 RpoB类型(推定的利福平敏感菌株)的利福平最低抑菌浓度(MIC)。一般来说,N531 RpoB类型的MIC水平(16 - 128微克/毫升)高于S531 RpoB类型(0 - 8微克/毫升)。为了确定N531 RpoB类型相对于利福平浓度的分离频率,我们对韩国土壤应用了涉及不同利福平浓度(0、20和100微克/毫升)的筛选方法。在较高的利福平浓度下观察到N531 RpoB类型的分离频率更高。此外,在本研究过程中,我们开发了一种等位基因特异性PCR方法来检测利福平抗性链霉菌菌株。我们的结果有力地表明,N531可能参与了链霉菌属菌株天然利福平抗性的主要机制。