Developing genetic transformation system in veillonellae

描述(申请人提供)：龋齿和牙周炎是人类最常见的两种疾病。虽然不会危及生命，但其发病率继续造成重大的社会和经济负担。这些疾病的多菌性使它们很难治愈，使用疫苗等传统手段进行预防更具挑战性。只有对牙科生物膜群落的内部工作及其与宿主的关系有了更深入的了解，整体或益生菌方法才能提供健康益处。韦洛氏菌一直被发现与龋齿和潜在的牙周炎有关，并被推测对牙齿生物膜的形成和发病有重要贡献。然而，由于这类细菌缺乏基因操纵系统，我们对它们在健康和疾病中的作用的了解充其量也是有限的。本项目旨在建立一套健壮的韦洛氏菌遗传转化系统。在第一次资助期间，我们成功地利用微小弧菌菌株PK1910获得了该属第一个真正的转化子。本项目通过提高转化效率、构建穿梭载体和创造转化受体品系，延续了前人的研究成果。这些研究的完成将建立一个强大的韦氏杆菌遗传转化系统，这将为研究人员提供必要的工具，在受控的实验室条件下研究韦氏杆菌的生物学、生理学、生态学和毒力。 公共卫生相关性：龋齿和牙周炎是人类最常见的两种疾病，分别影响大多数儿童和成年人，即使在美国这样的发达国家也是如此。这些疾病的多菌病原学使它们很难治愈，通过接种疫苗进行预防更具挑战性。基于了解牙菌斑中微生物生态的整体或益生菌方法可以为疾病预防和管理提供可行的选择。本项目旨在开发一种遗传学工具来研究牙菌斑中一组重要细菌(韦氏杆菌)的生物学、生理学、生态学和致病机理，从而为疾病预防提供新的方法。

DESCRIPTION (provided by applicant): Dental caries and periodontitis are two most prevalent diseases of mankind. Although not life threatening, their morbidity continues to imposed a significant social and economic burden. The polymicrobial nature of these diseases makes them difficult to cure, and even more challenging for prevention using conventional means such as vaccination. Holistic or probiotic approaches could offer health benefits only with a deeper understanding of the inner workings of the dental biofilm community as well as their relationships with the host. Veillonella species are consistently found to be associated with dental caries and potentially periodontitis, and are speculated to contribute significantly to dental biofilm development and pathogenesis. However, due to the lack of a genetic manipulation system in this group of bacteria, our knowledge about their role in health and disease is limited at best. This project aims to establish a robust Veillonella genetic transformation system. In the first grant period, we have succeeded in obtaining the first true transformant in this genus using V. parvula strain PK1910. The current project continues the previous studies by improving transformation efficiency, constructing shuttle vectors, and creating a transformation recipient strain. Completion of these studies will establish a robust genetic transformation system for veillonellae, which will provide researchers the tools required to study Veillonella biology, physiology, ecology, and virulence under controlled laboratory conditions. PUBLIC HEALTH RELEVANCE: Dental caries and periodontitis are two most prevalent diseases of mankind, affecting the majority of children and adults, respectively, even in developed countries like the U.S. The polymicrobial etiology of these diseases makes them difficult to cure, and even more challenging for prevention through vaccination. Holistic or probiotic approaches based on understanding the ecology of the microorganisms in the dental plaque could provide viable options for disease prevention and management. This project aims to develop a genetic tool to study the biology, physiology, ecology and pathogenesis of a group of important bacteria (veillonellae) in the dental plaque, so that novel approaches can be developed for disease prevention.