红色毛癣菌蛋白质组学研究
发布时间：2018-11-18 06:34
【摘要】： 红色毛癣菌(Trichophyton Rubrum)感染所致的浅部真菌病在世界各地广泛流行,红色毛癣菌感染难以治愈,且抗真菌药物停止治疗后频繁复发。本研究首次从整体水平对静止期红色毛癣菌分生孢子的蛋白质组进行全面的分析。 本研究首先构建了红色毛癣菌8个时相的cDNA文库,建立了EST编码蛋白数据库。然后采用液氮研磨、蜗牛酶消化后样品预分级,同时结合多种酶消化,酶切后得到的肽片段混合物通过二维液相色谱分离,采用鸟枪法对蛋白进行鉴定。结果获得5304个不同的肽段序列,总计1026个蛋白得到鉴定。我们建立蜗牛酶消化破壁,结合样品预分级,然后胰酶和蛋白酶K进行酶解的蛋白提取分级分离方法,可以显著提高鉴定的蛋白数量。 我们对所鉴定的1026个蛋白分别用NCBI non-redundant (NR)蛋白数据库, eukaryotic orthologous groups (KOG)数据库和gene ontology (GO)数据库进行检索,所鉴定的蛋白涵盖了所有主要的细胞器,如细胞核、线粒体、细胞膜、高尔基体、内质网,所鉴定的蛋白涉及到几乎所有的生物生化过程。本研究鉴定出红色毛癣菌分生孢子一些重要的蛋白,包括细胞壁结构蛋白、细胞壁生物合成相关蛋白,胞内蛋白质合成相关蛋白,静止期分生孢子代谢特征蛋白,信号传导蛋白以及环境压力耐受蛋白等等,并从蛋白分子水平上对其结构功能和机制进行了详细的讨论,红色毛癣菌分生孢子蛋白质组的分析结果证明了维持孢子的休眠状态是相当复杂和精细的过程,涉及到一系列的蛋白翻译、信号传导、环境压力耐受机制等,对将来真菌感染的预防、诊断,治疗和致病机制的研究有重要的提示作用。
[Abstract]:The superficial mycosis caused by (Trichophyton Rubrum) infection of Trichophyton rubrum is widespread all over the world. It is difficult to cure the infection of Trichophyton rubrum and recur frequently after the treatment of antifungal drugs. In this study, the proteome of conidia of Trichophyton rubrum at the whole level was analyzed for the first time. In this study, the cDNA library of 8 phases of Trichophyton rubrum was constructed, and the EST coding protein database was established. Then the samples were ground with liquid nitrogen, pre-graded by snail's enzyme digestion, and digested by many kinds of enzymes. The mixture of peptide fragments was separated by two-dimensional liquid chromatography, and the protein was identified by bird gun method. Results 5304 different peptide sequences were obtained and a total of 1026 proteins were identified. We established a method of protein extraction and fractionation of snail's enzyme digestion and broken wall, combined with sample pre-grading, and then enzymatic hydrolysis of trypsin and protease K. the number of identified proteins can be significantly increased. We searched the 1026 identified proteins using the NCBI non-redundant (NR) protein database, eukaryotic orthologous groups (KOG) database and the gene ontology (GO) database respectively. The identified proteins covered all major organelles such as nuclei and mitochondria. Membrane, Golgi body, endoplasmic reticulum, identified proteins involved in almost all biochemical processes. In this study, some important proteins in conidia of Trichophyton rubrum were identified, including cell wall structural protein, cell wall biosynthesis related protein, intracellular protein synthesis related protein, resting conidial metabolic characteristic protein. Signal transduction proteins and environmental pressure tolerance proteins are discussed in detail, and their structure, function and mechanism are discussed at the level of protein molecules. The analysis of conidial proteome of Trichophyton rubrum proved that maintaining the dormancy of spores is a very complicated and delicate process, involving a series of protein translation, signal transduction, environmental stress tolerance mechanism, etc. It has important implications for the prevention, diagnosis, treatment and pathogenesis of fungal infections in the future.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R379
[Abstract]:The superficial mycosis caused by (Trichophyton Rubrum) infection of Trichophyton rubrum is widespread all over the world. It is difficult to cure the infection of Trichophyton rubrum and recur frequently after the treatment of antifungal drugs. In this study, the proteome of conidia of Trichophyton rubrum at the whole level was analyzed for the first time. In this study, the cDNA library of 8 phases of Trichophyton rubrum was constructed, and the EST coding protein database was established. Then the samples were ground with liquid nitrogen, pre-graded by snail's enzyme digestion, and digested by many kinds of enzymes. The mixture of peptide fragments was separated by two-dimensional liquid chromatography, and the protein was identified by bird gun method. Results 5304 different peptide sequences were obtained and a total of 1026 proteins were identified. We established a method of protein extraction and fractionation of snail's enzyme digestion and broken wall, combined with sample pre-grading, and then enzymatic hydrolysis of trypsin and protease K. the number of identified proteins can be significantly increased. We searched the 1026 identified proteins using the NCBI non-redundant (NR) protein database, eukaryotic orthologous groups (KOG) database and the gene ontology (GO) database respectively. The identified proteins covered all major organelles such as nuclei and mitochondria. Membrane, Golgi body, endoplasmic reticulum, identified proteins involved in almost all biochemical processes. In this study, some important proteins in conidia of Trichophyton rubrum were identified, including cell wall structural protein, cell wall biosynthesis related protein, intracellular protein synthesis related protein, resting conidial metabolic characteristic protein. Signal transduction proteins and environmental pressure tolerance proteins are discussed in detail, and their structure, function and mechanism are discussed at the level of protein molecules. The analysis of conidial proteome of Trichophyton rubrum proved that maintaining the dormancy of spores is a very complicated and delicate process, involving a series of protein translation, signal transduction, environmental stress tolerance mechanism, etc. It has important implications for the prevention, diagnosis, treatment and pathogenesis of fungal infections in the future.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R379
【参考文献】
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1 李恒进,樊建峰,索继江,李若瑜,万U，
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