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所在平台: Udemy |
课程主页: https://www.udemy.com/course/bacteriophages-isolation-characterization-protocols/
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**课程名称：** 噬菌体分离与鉴定：实验方案 **课程概述：** 本课程聚焦于噬菌体（Bacteriophages）——专门感染细菌的病毒。在全球抗生素耐药性日益严峻的背景下，噬菌体作为一种有潜力的抗生素替代疗法，正受到研究和临床的关注。历史上，噬菌体疗法曾是治疗细菌感染的首选方法。 课程强调，并非所有噬菌体都适合用于治疗。例如，溶原性噬菌体（prophages）可能导致细菌细胞溶原化而非裂解，因此不应作为治疗选择。为了提供多样化的治疗方案，需要广泛的噬菌体资源。 尽管噬菌体本身未显示直接毒性，但研究其与哺乳动物细胞的相互作用至关重要。为确保结果的准确性，噬菌体制备物必须不含细菌细胞、毒素及其他可能干扰宿主反应的物质。 negativa染色结合电子显微镜是鉴定噬菌体的金标准。 基因组学的研究极大地深化了我们对噬菌体生物学的认识。通过对噬菌体进行全基因组测序，可以全面了解其生物学特性，并确认其生命周期。基因测序有助于深入理解噬菌体编码的蛋白质及生物分子（特别是裂解酶），这些物质在细菌细胞裂解及死亡过程中发挥关键作用。质谱技术则可用于鉴定噬菌体的结构蛋白。 作为生物防治剂使用裂解噬菌体，需要采用最适宜的标准化方法以确保应用安全。本课程旨在教授分子生物学技术，以支持噬菌体的分离和鉴定，从而促进后续研究与应用。 **教学重点：** * 噬菌体作为抗生素替代疗法的潜力 * 避免使用溶原性噬菌体 * 噬菌体制备物的纯化要求 * 噬菌体鉴定方法： negativa染色与电子显微镜 * 基因组学在噬菌体研究中的作用 * 噬菌体基因测序的意义：理解裂解机制 * 质谱在噬菌体结构蛋白鉴定中的应用 * 噬菌体作为生物防治剂的安全应用标准 * 分子生物学技术在噬菌体分离与鉴定中的实践
The world is on the cusp of a post-antibiotic period. A century ago, before the advent of antibiotics, bacteriophage therapy was the treatment of choice for bacterial infections. Although bacteriophages have yet to be approved as a treatment in Western medicine, researchers and clinicians have begun to anticipate phage therapy. Bacteriophages are viruses that depend on bacterial cell metabolism to multiply. They offer a promising alternative to the use of antibiotics and an excellent antibacterial option for combating multidrug resistance in bacteria. However, not every phage is suitable for phage therapy. In particular, prophages should not be used because they can lysogenize host cells instead of lysing them. To offer adequate therapeutic options for patients suffering from various infectious diseases, a wide selection of different phages is needed. While there is no evidence of direct toxicity induced by phage particles, it is crucial to study mammalian cell-phage interactions. This requires phage preparations to be free of bacterial cells, toxins and other compounds to avoid skewing host responses. Negative staining of purified viruses and electron microscopy remain the gold standard in the identification of bacteriophages. Interestingly, genomics has greatly changed our understanding of phage biology. Bacteriophage genome sequencing is essential to obtain a complete understanding of the bacteriophages' biology and to obtain confirmation of their lifestyle. Full genetic sequencing of bacteriophage will enable a better understanding of the phage-encoded proteins and biomolecules (especially phage lytic enzymes) involved in the process of bacterial cell lysis and death. Mass spectrometry can be used for the identification of phage structural proteins. The use of lytic phages as biocontrol agents requires the most appropriate and standard methods to ensure application safety. This course pursues methods in molecular biology for the isolation and characterization of phages to facilitate follow-up and implementation of work for other researchers.
