红色毛癣菌孢子EST构建及基因表达谱分析

发布时间：2018-10-12 20:27

【摘要】： 红色毛癣菌是重要的浅表感染真菌。世界范围内,60%的浅部真菌病由该菌引起。本文构建了红色毛癣菌BMU01672的cDNA文库,并进行大规模测序。通过聚类分析获得了10188条无冗余基因表达序列标签(Expression Sequence Tags, EST)。通过与现有数据库进行比较和利用COG软件对这些EST进行功能分类,明确了部分基因的功能以及代谢途径。为了进一步研究红色毛癣菌的的生理过程、致病机制和代谢特点,我们构建了包含10250条红色毛癣菌EST的cDNA芯片。并用此对红色毛癣菌的孢子萌发过程进行了研究。根据实验观察和文献报道,我们将红色毛癣菌孢子萌发过程分为16个时相,分别提取其mRNA作芯片杂交。在对所得数据进行筛选后,总共获得共1576个有效数据。对这1576个基因进行聚类分析,使用K-means聚类,最后选择的聚类clusters数目为5,经过SDSS 10.0统计软件验证,证明了聚类间的统计学意义。对红色毛癣菌在孢子生长过程中的基因表达变化和具体的生理进程之间的关系进行研究。根据GO数据库分类,对实验中所获得的差异基因进行了分类研究,以对不同功能的基因在孢子萌发时可能的作用进行分析。同时将这1576个ESTs与酵母基因数据库(the Saccharomyces Genome Database,SGD)进行了对比研究,通过比对同源基因在红色毛癣菌孢子生长过程中表达量的变化,分别对这些信号通路在红色毛癣菌孢子萌发中的作用进行了分析。
[Abstract]:Trichophyton rubrum is an important superficial infective fungus. Worldwide, 60% of superficial mycosis is caused by this fungus. The cDNA library of Trichophyton rubrum BMU01672 was constructed and sequenced on a large scale. 10188 non-redundant gene expression sequence tags (Expression Sequence Tags, EST). Were obtained by cluster analysis. By comparing with existing databases and using COG software to classify the functions of these EST, the functions and metabolic pathways of some genes were clarified. In order to further study the physiological process, pathogenic mechanism and metabolic characteristics of Trichophyton rubrum, we constructed a cDNA microarray containing 10250 EST of Trichophyton rubrum. The spore germination process of Trichophyton rubrum was studied. According to the experimental observation and literature reports, we divided the spore germination process of Trichophyton rubrum into 16 time phases and extracted its mRNA for microarray hybridization. After screening, a total of 1576 valid data were obtained. The 1576 genes were clustered by K-means, and the number of cluster clusters was 5, which was verified by SDSS 10.0 statistical software, which proved the statistical significance of clustering. The relationship between gene expression and physiological process of Trichophyton rubrum during spore growth was studied. According to the classification of GO database, the differential genes obtained in the experiment were classified and studied to analyze the possible role of genes with different functions in spores germination. At the same time, 1576 ESTs were compared with yeast gene database (the Saccharomyces Genome Database,SGD. By comparing the expression of homologous genes in the spore growth process of Trichophyton rubrum, we compared the expression levels of homologous genes in the spores of Trichophyton rubrum. The effects of these signaling pathways on spore germination of Trichophyton rubrum were analyzed.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2007
【分类号】：R346

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