具有抑菌活性的真菌的鉴定及其抑菌谱的初步研究
Abstract
In recent years, the development of new antimicrobial substances to replace the traditional antibiotics and fungicides has gradually become the focus of research. The topics to screen the antibacterial activity of strain as the starting point, isolated from a strain with good antibacterial activity of fungi, extracted the genome and PCR amplification of the internal transcribed spacer fragment. After purification were sequenced. Results by blast strain identification. In addition, Escherichia coli, Staphylococcus aureus and Bacillus subtilis were used as indicator bacteria, and the antibacterial spectrum of this strain was studied by Oxford cup method. The results of ITS identification showed that the bacterial strain was the result of the fermentation and metabolism of Escherichia coli, Staphylococcus aureus and Bacillus subtilis. This study is expected to provide reference for the further improvement of the antibacterial activity of this strain and the development of antibacterial products
毕业论文关键词：真菌; 抑菌活性; 鉴定; 抑菌谱
Keyword： Fungus；Antibacterial activity；Identification；Antimicrobial spectrum
目录
1.材料和方法 4
1.1 材料 4
1.1.1试验菌种 4
1.1.2仪器设备 5
1.1.3实验试剂 5
1.2方法 6
1.2.1 培养基的配置 6
1.3 菌种鉴定 7
1.3.1 基因组提取 7
1.3.2 PCR扩增 8
1.3.2.1 20ulPCR反应体系 8
1.3.2.2PCR反应引物序列 8
1.3.2.3 PCR反应条件 8
1.3.3 琼脂糖凝胶电泳 9
1.3.4 PCR产物纯化 9
1.3.5 ITS测序 9
1.3.6 序列比对 9
1.4抑菌谱研究 10
1.4.1指示菌菌液制备 10
1.4.2菌种发酵培养与代谢产物的收集 10
1.4.3牛津杯抑菌法 10
2.结果与讨论 11
2.1形态学观察 11
2.2 菌种发酵培养与代谢产物的收集 12
2.3 PCR扩增 13
2.4 ITS测序 13
2.5序列比对 14
2.6同源性比对 15
2.7抑菌谱的初步研究 16
3.结论 18
引言
现在世界上依然存在着三种急需解决的感染性疾病，依次是是获得性免疫缺陷综合症（HIV），乙肝和MRSA感染，在这些疾病中金黄色葡萄球菌是根源性的病原菌。专家开始不断的研究在动植物，微生物中不断筛选出能够对金黄色葡萄球菌具有抑菌活性的物质或者菌种。有相关研究报道有专家从深海微生物中分离出抗金黄色葡萄球菌的菌株12。从深海淤泥中分离菌株可行，但取样不方便，工作量大。查看相关文献发现最初始研究的专家通常是从土壤中分离出来菌株，进行抗金黄色葡萄球菌的研究。参考诸多文献决定本课题从土壤中取样分离菌株进行研究，土壤中取样具有取样方便，微生物种类多样的特点。