Mycobacteriophage as an emerging model organism

描述（由申请人提供）：分枝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等基本问题的首要模型系统。分枝噬菌体是感染分枝杆菌的病毒，易于分离、生长和基因组表征。已经对100多个分枝噬菌体基因组进行了测序——这是感染单一共同宿主的最大噬菌体群——但这只是全球1031个分枝噬菌体中占主导地位的一小部分。分枝噬菌体种类繁多，其中85%的基因功能未知。由于缺乏基因操作系统来进行基因敲除、插入和添加基因标签，阐明基因功能和揭示分枝噬菌体的生物学秘密是不可能的。这个问题被用于基因组操作的育种技术的发展所规避。这是第一次，开发一个简单和广泛适用的分枝噬菌体遗传系统是可能的。由于有近1万个基因，其中大部分功能未知，因此对这些工具的需求日益迫切。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，它目前正在爆炸式增长。首先是由HHMI科学教育联盟在美国各高校实施的噬菌体发现和基因组学，到2011年包括36个机构，75名教师和数百名学生研究人员，其中许多人需要扩展他们的研究以解决噬菌体生物学中的特定问题，并且需要这样做的工具。其次，新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，基因必要性确定，插入突变体和标记基因的创建，以及转座突变系统。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网站将为噬菌体基因组数据、资源传播、材料请求、建立合作和材料共享提供一个中心位置。

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.