【摘要】 【背景】金黄色葡萄球菌是重要的致病菌,其中促凝聚是重要的致病机制之一,可能存在新的基因参与其中。【目的】通过采用血浆凝集降低方法筛选及鉴定促凝相关基因。【方法】利用转座子随机插入突变技术建立金黄色葡萄球菌转座子突变文库,采用动态比浊及试管凝集技术筛选凝集能力降低的突变株;应用抗性标记挽救法鉴定突变基因并应用生物信息学预测基因的功能。【结果】通过观察血浆凝集能力降低共计筛选到突变菌株82个。鉴定其中的76个突变菌株的转座子插入位点,涉及基因13个,包括报道的与促凝集有关基因4个(占筛选基因的30.8%)。【结论】从金黄色葡萄球菌凝集能力降低筛选与促凝集可能有关的基因,为金黄色葡萄球菌促凝集基因的筛选提供了新策略,同时为了解该菌促凝集过程提供了候选基因。更多还原

【Abstract】 [Background] Staphylococcus aureus is an important pathogen, and procoagulation is one of the most important pathogenic mechanism, possibly novel genes in this approach. [Objective] To screen and identify procoagulant genes of Staphylococcus aureus by observing the decreased agglutination of colony phenotype. [Methods] Random mutagenesis library was constructed by using Tn917 transposon system, and mutants would be screened by combining dynamic turbidimetry and tube agglutination. The related genes were identified, and putative function of these genes was predicted through bioinformatics analysis. [Results] A total of 82 mutants were selected in a way of decreased agglutination phenotypes, and insertion sites of 76 mutants were successfully identified with 13 mutated genes, including 4 were reported in previous studies(which is 30.8% of these genes identified). [Conclusion] We acquired related genes based on decreased agglutination of mutants, and this was a new method of identifying potential genes involved in procoagulant ability, at the same time, new agglutination candidates obtained were reserved for further study.更多还原