小鼠AMcase基因克隆表达、蛋白纯化及抗真菌功能的初步研究

发布时间：2018-01-06 08:35

本文关键词：小鼠AMcase基因克隆表达、蛋白纯化及抗真菌功能的初步研究　出处：《第四军医大学》2006年硕士论文　论文类型：学位论文

【摘要】：近年来随着免疫受损人群的不断扩大，真菌性机会感染的临床地位显得越来越重要。现有的抗真菌药物在发挥治疗作用的同时，存在肝肾毒性、易产生耐药性和抗菌谱单一等问题。因此寻找能专一作用于真菌靶标的高效、安全、广谱的抗真菌药物已是研究抗真菌药物的重要方向。几丁质作为真菌细胞壁的主要结构成分，其代谢的重要性不言而喻。由于人体细胞没有细胞壁且人体不含有几丁质，，因此其特异性的水解酶—酸性哺乳动物几丁质酶(AMCase)可能是安全、有效抗真菌剂的理想候选对象。课题目的：克隆小鼠AMCase基因，应用生物信息学技术从理论上分析AMCase蛋白潜在的抗真菌功能。表达纯化小鼠AMCase蛋白，并同时从小鼠胃肠组织中分离提纯天然AMCase蛋白，通过体外抗真菌试验研究，明确AMCase对红色毛癣菌、白念珠菌和烟曲霉3种病原真菌的抗真菌作用，为开发抗真菌生物治疗的绿色方案提供新的策略。
[Abstract]:In immunocompromised patients, the clinical status of opportunistic fungal infection is becoming more and more important. Antifungal agents in the therapeutic effects at the same time, there are liver and kidney toxicity, drug resistance and single antimicrobial spectrum. So looking for specific antifungal targets for efficient, safe, broad-spectrum anti has antifungal drugs is an important research direction of antifungal drugs.
Chitin as the main structure of fungal cell wall components, its importance is self-evident. The metabolism of human cells without cell wall and the body does not contain chitin, therefore the hydrolase specific acidic mammalian chitinase (AMCase) may be safe, ideal candidates for effective antifungal agents.
The purpose of the project:
Cloning of mouse AMCase gene by bioinformatics technology from the theoretical analysis of antifungal proteins AMCase potential. Expression and purification of mouse AMCase protein, and also from the gastrointestinal tissues isolated from mice with natural AMCase protein purified by in vitro experimental study on anti fungal, clear AMCase of Trichophyton rubrum, Candida albicans and Aspergillus fumigatus 3 pathogens fungi antifungal effects and provide a new strategy for green solutions for the development of anti fungal biological treatment.

【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346

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