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首页> 外文会议>12th Symposium on Bioluminescence and Chemiluminescence, Apr 5-9, 2002, Robinson College, University of Cambridge, UK >A NOVEL DUAL REPORTER ASSAY FOR STUDYING INTRACELLULAR BACTERIAL PATHOGENS
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A NOVEL DUAL REPORTER ASSAY FOR STUDYING INTRACELLULAR BACTERIAL PATHOGENS

机译：研究细胞内细菌性病原菌的新型双重报告检测

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Staphylococcus aureus is a versatile pathogen, whose infections, due to the increasing emergence of antibiotic resistant strains, are causing considerable alarm within the medical community. The expression of many S. aureus virulence factors is under the control of the agr quorum sensing system. A model was proposed for the function of agr mediated quorum sensing in staphylococcal invasion of cells, which suggested that arg related products were necessary for escape from the endosome and subsequent release and growth in the cytoplasm. It was proposed that in an extracellular environment, agr was down-regulated due to dilution of the agr peptide in the surrounding fluids. Upon uptake, the agr peptide concentrations increased rapidly when the bacteria were confined within the endosome, and that lysis occurred due to the induction of agr related exoproteins. Our previous studies with S. aureus RN6390 support the hypothesis that agr expression precedes host endosomal lysis. Recent studies have shown that S. aureus strains derived from the NCTC 8335 lineage, (e.g. RN6390, 8325-4) carry a deletion in rsbU. As rsbU encodes a positive regulator for the activation of the stress response sigma factor SigB, the deletion results in cells that are phenotypically sigB defective. If internalisation induces a stress response in S. aureus, the defect in rsbU is expected to affect the level of agr expression and change the internalisation capabilities of the organism. A recent study indicates that differences in the regulation of sarA and agr exist within different strains of S. aureus, but it is uncertain if this would affect virulence. As our previous experiments were performed with S. aureus RN6390, the data may be at variance with that obtained with 'wild type' S. aureus. This study investigates the possible similarities or differences of S. aureus rsbU~+ and rsbU~- strains using a novel GFP-Lux dual reporter system and high throughput microtitre plate assay.
机译：金黄色葡萄球菌是一种多功能病原体，由于抗生素耐药菌株的不断出现，其感染已引起医学界的广泛关注。许多金黄色葡萄球菌毒力因子的表达在农业群体感应系统的控制下。提出了一种模型，用于在农杆菌介导的细胞中由agr介导的群体感应功能，这表明arg相关产物对于从内体逃逸以及随后在细胞质中的释放和生长是必需的。有人提出，在细胞外环境中，由于周围液体中的agr肽稀释，导致agr下调。摄取后，当细菌被限制在内体中时，agr肽浓度会迅速增加，并且裂解是由于agr相关外蛋白的诱导而发生的。我们先前对金黄色葡萄球菌RN6390的研究支持agr表达先于宿主内体裂解的假说。最近的研究表明，源自NCTC 8335谱系的金黄色葡萄球菌菌株（例如RN6390、8325-4）在rsbU中带有缺失。由于rsbU编码用于激活应激反应sigmaB因子SigB的正调节剂，因此删除会导致表型为sigB缺陷的细胞。如果内部化在金黄色葡萄球菌中引起应激反应，则rsbU中的缺陷有望影响agr表达水平并改变生物体的内部化能力。最近的一项研究表明，在不同菌株的金黄色葡萄球菌中存在sarA和agr的调控差异，但是尚不确定这是否会影响毒力。由于我们先前的实验是用金黄色葡萄球菌RN6390进行的，因此该数据可能与“野生型”金黄色葡萄球菌获得的数据不一致。本研究使用新型GFP-Lux双报告系统和高通量微量滴定板分析法调查金黄色葡萄球菌rsbU〜+和rsbU〜-菌株的可能相似性或差异性。

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