Development of Genetic techniques in Chlamydia

描述(申请人提供)：沙眼衣原体可导致人类眼睛和生殖器感染，可导致失明和不孕。肺炎衣原体感染是肺炎的原因之一，也是慢性阻塞性肺疾病、哮喘和动脉粥样硬化的危险因素。尽管衣原体对人类健康有巨大影响，但人们对衣原体致病的遗传学知之甚少，因为不存在在这些生物中创造靶向突变的基因技术。因此，迫切需要开发基因系统来操纵衣原体的染色体。我们已经基于pCHL1(沙眼衣原体中发现的自然质粒)创建了质粒，这将为基因中断系统提供基础，以启动对这一难以捉摸的细菌群体的遗传操作。我们将以此为基础发展衣原体的遗传操作技术。我们将首先优化转化条件，以分离携带该质粒的沙眼衣原体和鼠疫杆菌。然后，我们将调整该质粒，将第二组内含子送入衣原体，以促进位点特异性基因失活。衣原体的成功转化和基因修饰将推动对这一重要人类病原体的研究，并为开发针对这些细菌的新疗法和疫苗奠定基础。

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.