Development of Genetic techniques in Chlamydia

衣原体遗传技术的发展

• 批准号: 8470126
• 负责人: Karl E Klose
• 金额: $ 7.35万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-16 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8470126
• 关键词: Asthma; Atherosclerosis; Bacteria; Blindness; Budgets; Catalytic RNA; Categories; Cell Separation; Chlamydia; Chlamydia trachomatis; Chlamydophila pneumoniae; Chromosomes; Chronic Obstructive Airway Disease; Communicable Diseases; Development; Disease; Ensure; Funding; Funding Mechanisms; Gene Silencing; Genes; Genetic; Genetic Techniques; Genetic Transformation; Growth; Health; Human; Infection; Infertility; Introns; Laboratories; Lead; Modification; Nature; Organism; Pathogenesis; Plaque Assay; Plasmids; Pneumonia; Recombinants; Relative (related person); Reporting; Risk Factors; Series; Sexually Transmitted Diseases; Site; Sterility; System; Techniques; base; cost; design; experience; falls; field study; genetic manipulation; genital infection; high reward; high risk; mutant; novel; novel therapeutics; novel vaccines; pathogen; promoter; research study; vector

DESCRIPTION (provided by applicant): Chlamydia trachomatis causes ocular and genital infections in humans, which can lead to blindness and infertility. Chlamydia pneumoniae infections are a cause of pneumonia and a risk factor for chronic obstructive pulmonary disease, asthma, and atherosclerosis. Despite their immense impact on human health, relatively little is known about the genetics of Chlamydia pathogenesis, because no genetic techniques exist to create targeted mutants in these organisms. Thus there is a pressing need for the development of genetic systems to manipulate the chromosome of Chlamydia. We have created plasmids based on pCHL1, the natural plasmid found in C. trachomatis, and this will provide the basis of a gene disruption system to initiate genetic manipulation of this elusive group of bacteria. We will use this plasmid as the basis for the development of genetic manipulation techniques in Chlamydiae. We will first optimize transformation conditions to isolate both C. trachomatis and C. muridarum carrying the plasmid. We will then adapt the plasmid to deliver a Group II intron into Chlamydia, to facilitate site-specific gene inactivation. Successful transformation and genetic modification of Chlamydia species will propel the study of this important group of human pathogens forward, and allow for the development of novel therapeutics and vaccines against these bacteria.

描述（由申请人提供）：沙眼衣原体会引起人类眼部和生殖器感染，从而导致失明和不育。肺炎衣原体感染是肺炎的原因之一，也是慢性阻塞性肺病、哮喘和动脉粥样硬化的危险因素。尽管衣原体对人类健康影响巨大，但人们对衣原体发病机制的遗传学知之甚少，因为没有遗传技术可以在这些生物体中产生有针对性的突变体。因此，迫切需要开发遗传系统来操纵衣原体的染色体。我们基于 pCHL1（沙眼衣原体中发现的天然质粒）创建了质粒，这将为基因破坏系统提供基础，以启动对这一难以捉摸的细菌群的基因操作。我们将使用该质粒作为衣原体遗传操作技术开发的基础。我们将首先优化转化条件以分离携带质粒的沙眼衣原体和鼠衣原体。然后，我们将调整该质粒，将 II 组内含子传递到衣原体中，以促进位点特异性基因失活。衣原体物种的成功转化和基因改造将推动对这一重要人类病原体的研究向前发展，并允许开发针对这些细菌的新型疗法和疫苗。