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【目的】鉴定白念珠菌肌醇多磷酸激酶Kcs1蛋白,并探索Kcs1在该病原菌细胞自噬、菌丝发育及致病过程中的功能。【方法】采用二步PCR介导的同源重组方法,构建白念珠菌KCS1基因缺失菌株kcs1Δ/Δ及回补菌株KCS1c;采用氮饥饿敏感性测定及GFP-Atg8自噬报告系统,测定KCS1缺失对白念珠菌自噬过程的影响;采用菌丝诱导培养,测定KCS1缺失对白念珠菌菌丝发育能力的影响;采用巨噬细胞模型及小鼠系统性感染模型,分析KCS1缺失对白念珠菌感染宿主能力的影响。【结果】KCS1缺失造成白念珠菌氮饥饿耐受能力降低,氮饥饿条件下自噬相关蛋白Atg8的降解及转运水平下降,菌丝发育变缓,对巨噬细胞耐受及损伤能力减弱,但不影响菌株的小鼠系统性感染能力。【结论】白念珠菌肌醇多磷酸激酶Kcs1在细胞自噬、菌丝发育、与巨噬细胞相互作用等方面发挥重要作用。
【Objective】 To identify Candida albicans inositol polyphosphate kinase Kcs1 protein and to explore the function of Kcs1 in autophagy, mycelium development and pathogenicity in this pathogen. 【Method】 KCS1Δ / Δ and KCS1c strains were constructed by two-step PCR-mediated homologous recombination. KCS1 deletion was determined by nitrogen starvation sensitivity assay and GFP-Atg8 autophagy reporter system Candida albicans autophagy process; using mycelium-induced culture to determine KCS1 deletion on Candida albicans mycelium development; macrophage model and mouse systemic infection model KCS1 deletion of Candida albicans infection host capacity Impact. 【Result】 The results showed that the deletion of KCS1 led to the decrease of tolerance to Candida albicans nitrogen starvation. The degradation and transport of autophagy-related protein Atg8 decreased under nitrogen starvation, the mycelium developed slowly and its ability of tolerance and injury to macrophages decreased. However, Does not affect the ability of strains of systemic infection in mice. 【Conclusion】 Candida albicans inositol polyphosphate kinase Kcs1 plays an important role in autophagy, mycelium development and interaction with macrophages.