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

模式生物天蓝色链霉菌的系统诱变：完成研究界的重要资源

• 批准号: BB/E019242/1
• 负责人: Paul Dyson
• 金额: $ 27.85万
• 依托单位: Swansea University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE019242%2F1
• 关键词: Systematic; mutagenesis; model; organism; Streptomyces

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 A critical aspect of understanding gene function is the creation of mutants in which a specific gene is no longer active.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.

链霉菌是一种具有重要经济意义的迷人土壤细菌，因为它们生产人类已知抗生素的 70%。对于细菌来说不同寻常的是，它们表现出复杂的分化生命周期，这对抗生素的生产具有影响。由于其重要性，BBSRC 和 Wellcome Trust 资助了模式物种 S. coelicolor 的完整基因组测序。因此，我们知道这种细菌拥有大约 8000 个基因。然而，迄今为止我们只了解其中一小部分基因的生物学功能。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节的复杂性。此外，天蓝色链球菌是一种遗传上易于处理的模型，可用于了解其与结核分枝杆菌等重要人类病原体共享的保守过程。了解基因功能的一个关键方面是创建突变体，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，可以了解相关基因的功能。拟议的研究旨在完成多功能资源的构建，目前世界各地的研究人员正在使用该资源在天蓝色链球菌中创建特定的基因突变。该资源是通过将“跳跃基因”或转座子插入到每个天蓝色链球菌基因的 DNA 序列中而制成的。这些插入是在粘粒文库中克隆的基因拷贝中进行的：该文库用于获得天蓝色链球菌基因组序列。因此，对于每个携带大约 30 个连续基因的原始粘粒，我们构建了一个由单独粘粒组成的子文库，每个粘粒在每个基因中携带单独的插入。通过获得这些突变粘粒，研究人员可以轻松构建目标基因有缺陷的天蓝色链球菌突变体。任何研究人员都可以通过在线数据库获取有关系统粘粒诱变计划的信息，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，我们自己将利用该资源更详细地研究链霉菌如何适应自然环境中不同的水分含量。这本身就会影响生命周期和抗生素的生产。

项目成果

Reference Module in Biomedical Sciences

生物医学科学参考模块

• DOI: 10.1016/b978-0-12-801238-3.02306-0
• 发表时间: 2014
• 作者: Dyson P

The evolution of an osmotically inducible dps in the genus Streptomyces.

• DOI: 10.1371/journal.pone.0060772
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Facey PD;Hitchings MD;Williams JS;Skibinski DO;Dyson PJ;Del Sol R
• 通讯作者: Del Sol R

Cell-Biological Studies of Osmotic Shock Response in Streptomyces spp.

• DOI: 10.1128/jb.00465-16
• 发表时间: 2017-01-01
• 期刊: JOURNAL OF BACTERIOLOGY
• 影响因子: 3.2
• 作者: Fuchino, Katsuya;Flardh, Klas;Ausmees, Nora
• 通讯作者: Ausmees, Nora

The dpsA gene of Streptomyces coelicolor: induction of expression from a single promoter in response to environmental stress or during development.

• DOI: 10.1371/journal.pone.0025593
• 发表时间: 2011
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Facey PD;Sevcikova B;Novakova R;Hitchings MD;Crack JC;Kormanec J;Dyson PJ;Del Sol R
• 通讯作者: Del Sol R

A heterodimer of EsxA and EsxB is involved in sporulation and is secreted by a type VII secretion system in Streptomyces coelicolor.

• DOI: 10.1099/mic.0.037069-0
• 发表时间: 2010
• 期刊: Microbiology
• 影响因子: 1.5
• 作者: Sandra Akpe San Roman;P. Facey;L. Fernández-Martínez;C. Rodríguez;C. Vallín;R. del Sol;P. Dyson