Development of Genetic Tools for Clostridium botulinum

肉毒梭菌遗传工具的开发

• 批准号: 6759627
• 负责人: Eric A. Johnson
• 金额: $ 25.69万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6759627
• 关键词: Clostridium botulinum; RNA; antisense nucleic acid; bacterial genetics; biotechnology; bioterrorism /chemical warfare; gene expression; gene mutation; gene targeting; genetic manipulation; mutant; technology /technique development; transfection /expression vector; transposon /insertion element

DESCRIPTION (provided by applicant): Despite the importance of Clostridium botulinum as a human and animal pathogen, and its potential role in bioterrorism, the organism remains extremely poorly characterized with regard to cellular processes and their role in pathogenesis. This is almost entirely a consequence of ineffective genetic systems, and in particular integrational tools. This inability to easily generate stable mutants (directed or random) is common to all clostridial species, and severely hinders the ability of the scientific community at large to fully exploit clostridial genome information in hypothesis driven research. The goal of this project is specifically to develop the necessary enabling technology to overcome this impediment. The aims are: (1) to develop integrational vectors to introduce targeted mutations in genes of C. botulinum; (2) to identify effective transposons for random mutagenesis and (3) to assess antisense RNA strategies for modulation and analysis of C. botulinum gene expression. These aims will be accomplished through the complementary knowledge and expertise of a USA and UK laboratory. The developed genetic tools will pave the way for in depth analysis of not only the C. botulinum genome, but other clostridial genomes too. Their future deployment will, for example, lead to the elucidation of physiological factors that control growth and toxin production, pathogenesis, and developmental processes unique to clostridia and related organisms including endospore formation, resistance, germination and outgrowth.

描述（由申请人提供）：尽管肉毒梭菌作为人类和动物病原体的重要性及其在生物恐怖主义中的潜在作用，但在细胞过程及其在发病机理中的作用方面，生物体仍然非常差。这几乎完全是无效的遗传系统，特别是集成工具的结果。这种无法轻松产生稳定的突变体（定向或随机）是所有梭形物种共有的，并且严重阻碍了科学界在整个假设驱动的研究中完全利用梭菌基因组信息的能力。该项目的目的是专门开发必要的启用技术来克服这一障碍。目的是：（1）开发综合向量以在肉毒杆菌基因中引入靶向突变； （2）确定有效的转座子，用于随机诱变和（3）评估肉毒杆菌基因表达的调节和分析的反义RNA策略。这些目标将通过美国和英国实验室的互补知识和专业知识来实现​​。开发的遗传工具将不仅为肉毒杆菌基因组，其他梭状芽胞杆菌基因组进行深入分析铺平道路。例如，他们的未来部署将导致阐明生长和毒素产生，发病机理和发育过程的生理因素，以及相关生物所独有的，包括内孢子形成，耐药性，发芽，发芽和产物。