Mycobacteriophage as an emerging model organism

描述（由申请人提供）：分枝杆菌噬菌体正在成为理解基因表达、蛋白质结构和功能以及基因组进化中基本问题的首要模型系统。分枝杆菌噬菌体是感染分枝杆菌的病毒，并且易于分离、生长和基因组表征。超过100个分枝杆菌噬菌体基因组已被测序-感染单一共同宿主的最大的细菌群-但占全球1031个粒子的一小部分，这些粒子主宰着生物圈。分枝杆菌噬菌体具有惊人的多样性，其85%的基因功能未知。由于缺乏基因敲除、插入和添加基因标签的遗传操作系统，阐明基因功能和揭开分枝杆菌噬菌体的生物学秘密是不可能的。随着用于基因组操作的BRED技术的发展，这个问题被规避了。第一次，一个简单的和广泛适用的分枝杆菌噬菌体遗传系统的发展是可能的。由于有近10，000个基因，其中大部分功能未知，因此对这些工具的需求非常迫切。虽然分枝杆菌噬菌体研究人员的数量一直相当有限，但由于两个主要事件，目前正在爆炸。首先是由HHMI科学教育联盟在美国的学院和大学中实施分枝杆菌噬菌体发现和基因组学，到2011年将纳入36个机构，75名教师和数百名学生研究人员，许多人需要扩展他们的研究以解决分枝杆菌噬菌体生物学中的特定问题，并需要工具来做到这一点。其次，新的BRED技术的简单性使其具有普遍适用性。我们将开发的遗传工具将能够构建简单的无标记基因敲除、基因必要性测定、插入突变体和标记基因的创建以及转座诱变系统。我们还将生成包括噬菌体特异性引物、质粒载体和转录组数据在内的资源。phagesdb.org网站将为分枝杆菌噬菌体基因组数据、资源传播、材料请求、合作建立和材料共享提供一个中心位置。 公共卫生相关性：分枝杆菌噬菌体为基因功能、表达和调控以及理解基因组进化机制提供了新的范例。除了作为一种简单而强大的模式生物外，它们还为了解其宿主结核分枝杆菌提供了强大的工具，结核分枝杆菌比任何其他单一的传染性病原体都要杀死更多的人。

DESCRIPTION (provided by applicant): Mycobacteriophages are emerging as a premiere model system for understanding fundamental questions In gene expression, protein structure and function, and genome evolution. Mycobacteriophages are viruses that infect mycobacteria, and are easy to Isolate, grow, and genomically characterize. Over 100 mycobacteriophage genomes have been sequenced - the largest group of phages infecting a single common host - but a tiny fraction of the global population of 1031 particles dominating the biosphere. Mycobacteriophages are amazingly diverse and 85% of their genes are of unknown function. Elucidating gene functions and unraveling the biological secrets of the mycobacteriophages has not been possible for lack of a system for genetic manipulations to make gene knockouts, insertions, and to add gene tags. This problem was circumvented with development of BRED technology for genome manipulation. For the first time, development of a facile and broadly applicable mycobacteriophage genetic system is possible. With nearly 10,000 genes, mostly of unknown function, there is an emerging and critical need for these tools. While the number of mycobacteriophage researchers has been fairly modest, it is currently exploding because of two main events. First is the implementation of mycobacteriophage discovery and genomics by the HHMI Science Education Alliance in colleges and universities across the US, with inclusion by 2011 of 36 institutions, 75 faculty, and many hundreds of student researchers, many who need to extend their investigations to address specific questions in mycobacteriophage biology and need the tools to do so. Secondly, the simplicity of the new BRED technology makes it generally applicable. The genetic tools we will develop will enable construction of simple unmarked gene knockouts, gene essentiality determination, creation of insertion mutants and tagged genes, and s transposition mutagenesis systems. We will also generate resources including phage-specific primers, plasmid vectors, and transcriptome data. The phagesdb.org web site will provide a central location for mycobacteriophage genome data, dissemination of resources, material requests, establishment of collaborations, and sharing of materials. PUBLIC HEALTH RELEVANCE: Mycobacteriophages offer new paradigms for gene function, expression and regulation, as well as understanding the mechanisms of genome evolution. In addition to being a simple and powerful model organism, they also provide powerful tools for understanding their host, Mycobacterium tuberculosis, which kills more people than any other single infectious agent.