Development of Genetic techniques in Chlamydia

描述（由申请人提供）： 沙眼衣原体引起人类眼部和生殖器感染，可导致失明和不育。肺炎衣原体感染是肺炎的一个原因，也是慢性阻塞性肺病、哮喘和动脉粥样硬化的一个危险因素。尽管它们对人类健康有巨大的影响，但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靶向突变体。因此，迫切需要开发遗传系统来操纵衣原体的染色体。我们已经创建了基于pCHL1的质粒，pCHL1是在C.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遗传操作。我们将使用这种质粒作为衣原体遗传操作技术发展的基础。我们将首先优化转化条件以分离两种C. trachomatis和C.携带有质粒的鼠伤寒沙门氏菌然后，我们将调整质粒，将第二组内含子导入衣原体，以促进位点特异性基因失活。成功的转化和基因修饰衣原体物种将推动这一重要的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疗法和疫苗。 公共卫生相关性：衣原体属这些疾病对人类健康造成巨大的负面影响，包括不育和失明。这些研究旨在开发衣原体属的遗传技术，这将有助于更好地了解疾病，并导致新的治疗和疗法。

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. PUBLIC HEALTH RELEVANCE: Chlamydia spp. cause a number of human infectious diseases, most notably sexually transmitted disease and ocular disease, which have a huge negative impact on human health that includes sterility and blindness. These studies are aimed at developing genetic techniques for Chlamydia spp, which will allow for a better understanding of disease and lead to novel cures and therapies.