Systematic mutagenesis of the model organism Streptomyces coelicolor: completion of an essential resource for the research community

链霉菌是具有重要经济意义的迷人土壤细菌，因为它们产生人类已知的70%的抗生素。与细菌不同的是，它们表现出复杂的分化生命周期，这对抗生素生产有影响。由于其重要性，BBSRC和Wellcome Trust资助了模式物种S的完整基因组测序。天蓝色。因此，我们知道这种细菌拥有大约8000个基因。然而，到目前为止，我们只了解这些基因中一小部分的生物学功能。了解基因功能很重要，因为它使我们能够更多地了解链霉菌的生物学，特别是抗生素生产和分化调控的复杂性。此外，S. coelicolor是一种遗传学上易于处理的模型，以了解它与重要的人类病原体如结核分枝杆菌共享的保守过程。理解基因功能的一个关键方面是创造突变体，其中特定基因不再活跃。通过比较突变体与非突变体的特征，可以了解所讨论的基因的功能。拟议的研究旨在完成一个多功能资源的构建，该资源目前正被世界各地的研究人员用于在S.天蓝色。这种资源是通过将一个“跳跃基因”或转座子插入到每个S的DNA序列中而获得的。coelicolor基因这些插入是在粘粒文库中克隆的基因拷贝中进行的：该文库用于获得S。腔棘鱼基因组序列因此，对于每一个携带大约30个连续基因的原始粘粒，我们构建了一个由每个基因中携带单独插入的单独粘粒组成的子文库。有了这些突变的互补链，研究人员可以很容易地构建一个S。在感兴趣的基因中有缺陷的天蓝色突变体。任何研究人员都可以通过在线数据库获得系统性粘粒诱变程序的信息，使他们能够选择和订购特定的突变体粘粒供自己使用。此外，我们将利用这些资源更详细地研究链霉菌如何适应其自然环境中不同的水分含量。这本身就影响了生命周期和抗生素生产。

Streptomyces are fascinating soil bacteria of major economic importance as they produce 70% of antibiotics known to man. Unusually for bacteria, they exhibit a complex differentiating life-cycle that has implications for antibiotic production. Due to their importance, the BBSRC and Wellcome Trust funded the complete genome sequencing of the model species S. coelicolor. As a consequence, we know this bacterium possesses about 8000 genes. However, as yet we only understand the biological function of a small proportion of these genes. Understanding gene function is important as it enables us to learn more about the biology of Streptomyces and, in particular, the complexities of the regulation of antibiotic production and differentiation. In addition, S. coelicolor is a genetically tractable model to understand conserved processes it shares with important human pathogens such as Mycobacterium tuberculosis. A critical aspect of understanding gene function is the creation of mutants in which a specific gene is no longer active. By comparing characteristics of the mutant with those of a non-mutant, it is possible to learn about the function of the gene in question. The proposed research intends to complete the construction of a versatile resource that is currently being used by researchers worldwide to create specific gene mutations in S. coelicolor. This resource is made by insertion of a 'jumping gene' or transposon into the DNA sequence of each S. coelicolor gene. These insertions are made in gene copies cloned in a cosmid library: this library was used to obtain the S. coelicolor genome sequence. So for each original cosmid carrying approximately 30 contiguous genes, we construct a sub-library consisting of individual cosmids each carrying an individual insertion in each gene. With access to these mutant cosmids, researchers can easily construct a S. coelicolor mutant defective in a gene of interest. Information on the systematic cosmid mutagenesis programme is accessible to any researcher via an online database, allowing them to select and order specific mutant cosmids for their own use. In addition, we will ourselves use the resource to investigate in more detail how Streptomyces adapt to the varying moisture content of their natural environment. This itself impacts on the life-cycle and antibiotic production.