Steinhuber Andrea, Landmann Regine, Goerke Christiane, Wolz Christiane, Flückiger Ursula
Division of Infectious Diseases & Hospital Epidemiology, University Hospital Basel, Petersgraben 4, CH-4031 Basel, Switzerland.
Int J Med Microbiol. 2008 Oct;298(7-8):599-605. doi: 10.1016/j.ijmm.2007.09.008. Epub 2008 Mar 7.
Alpha-toxin (Hla, encoded by hla) is a major virulence factor of Staphylococcus aureus. The activity of the hla promoter was analyzed using luxABCDE on an integration vector. The phla-lux construct was introduced in S. aureus Newman and its isogenic sae and sigB regulator mutants. Promoter activity was monitored by bioluminescence in vitro and in the murine tissue-cage model. Hla promoter activity could be followed in real time at repeated time points of infection. The activation of hla in the sigB-deficient strain and the repression to background levels in a sae-deficient strain relative to hla expression in the wild type could be demonstrated in vivo. Subinhibitory concentrations of teicoplanin, imipenem and ciprofloxacin enhanced hla promoter activity in vitro whereas clindamycin and rifampicin did not. Our approach proved to be rapid and adequate to study promoter activity in vitro and in vivo under conditions where high bacterial numbers are reached.
α-毒素（由hla编码的Hla）是金黄色葡萄球菌的一种主要毒力因子。使用整合载体上的luxABCDE分析hla启动子的活性。将phla-lux构建体导入金黄色葡萄球菌Newman及其同基因的sae和sigB调节突变体中。通过体外和小鼠组织笼模型中的生物发光监测启动子活性。在感染的重复时间点可以实时跟踪Hla启动子活性。在体内可以证明，与野生型中的hla表达相比，sigB缺陷菌株中hla的激活以及sae缺陷菌株中hla表达被抑制至背景水平。替考拉宁、亚胺培南和环丙沙星的亚抑制浓度在体外增强了hla启动子活性，而克林霉素和利福平则没有。我们的方法被证明是快速且适用于在达到高细菌数量的条件下研究体外和体内启动子活性的。
