论文部分内容阅读
红色毛癣菌是最常见的人类浅部真菌病病原体之一,可引起皮肤和指甲慢性感染.为研究抗真菌药物作用下特异表达的基因,用cDNA芯片技术分析了5-氟胞嘧啶作用下的红色毛癣菌表达谱.从红色毛癣菌表达序列标签文库中挑选基因制备cDNA芯片,测定5-氟胞嘧啶的最低抑菌浓度(minimum inhibitory concentration,MIC),进行芯片杂交和数据分析,选取有代表性的7个基因用实时定量反转录多聚酶链反应技术(real-time reverser transcription polymerase chain reaction,Real-timeRT-PCR)进行验证.数据分析表明,有474个基因发生了差异性表达,196个基因表达上调,278个基因表达下调.核酸代谢途径中的CDC21,核酸转录因子E2F1,RNA代谢途径中的SGN1,RIM4和NOP1等基因出现明显下调表达.其他参与了信号转导、分子伴侣活性、离子转运、次级代谢途径、氨基酸转运、脂质转运和潜在耐药机制等作用途径的基因表达也发生相应改变.所选基因的Real-timeRT-PCR结果验证了芯片结果的可信性.本研究得到了第1个5-氟胞嘧啶作用下的红色毛癣菌表达谱,为进一步研究5-氟胞嘧啶的药物作用机制和耐药机理提供了有价值的信息.
Trichophyton rubrum is one of the most common human pathogens of shallow mycosis and can cause chronic infection of the skin and nails. In order to study the genes specifically expressed under the action of antifungal agents, the effect of 5-fluorocytosine Of Trichophyton rubrum expression profiles were selected from the cDNA library of Trichophyton rubrum expression sequence tagging gene to prepare a minimum inhibitory concentration (MIC) of 5-fluorocytosine for chip hybridization and data analysis, Seven representative genes were selected for validation by real-time RT-PCR.The data analysis showed that 474 genes were differentially expressed , 196 genes were up-regulated and 278 genes were down-regulated.CDC21, E2F1 and SGN1, RIM4 and NOP1 in RNA metabolic pathway were significantly down-regulated.The other involved in signal transduction, Gene expression of pathways of action such as chaperone activity, ion transport, secondary metabolic pathways, amino acid transport, lipid transport and potential drug resistance mechanisms The results of real-time RT-PCR of the selected genes validated the authenticity of the chip results.In this study, we obtained the expression profile of Trichophyton rubrum under the first 5-fluorocytosine, Flucytosine drug mechanism of action and resistance mechanisms provide valuable information.