Characterization of Culturable and Nonculturable Bacteria: A 16S rRNA Sequence Approach

本项目将利用分子生物学的工具， 长期存在的生物海洋学问题。 其主要目标是 了解哪种细菌在天然益生菌中占主导地位 种群，（B）产生异养菌纯培养物库 寡养型，和（c）确定哪些，如果有的话，文化是 代表自然种群的主要组成部分。直到 现在，大多数自然产生的细菌还没有被 遗传特征（或“鉴定”），因为它们具有 抵制试图培养他们，排除传统的 识别.该项目将规避问题， 最先进的RNA测序和DNA克隆技术 来表征微生物。这样的工作现在可以在新鲜的 收集材料而不培养微生物。这 项目将提供（1）明确识别 占主导地位的海洋细菌，在某种程度上适合比较物种 可培养和不可培养群体的组成，（2） 选择与那些最密切相关的分离物 占主导地位的“真实的”海洋种群，适合实验室研究，（3） 分子探针的发展，以确定人口分布， 在其他地区，从而衡量内部和之间的多样性 广泛不同的环境，（4）这些探头的可能用途 用于单细胞杂交研究， 鉴定保存样品中的微生物， 海洋环境

This project will use tools of molecular biology to approach a long-standing biological oceanographic problem. Its major goals are to (a) learn what kinds of bacteria dominate natural planktonic populations, (b) generate a bank of pure cultures of heterotrophic oligotrophs, and (c) determine which, if any, cultures are representative of major components of the natural populations. Until now, most of the naturally-occurring bacteria have not been phylogenetically characterized (or "identified") because they have resisted attempts to culture them, precluding conventional identification. This project will circumvent the problem with state-of-the-art techniques of RNA sequencing, and DNA cloning to characterize microorganisms. Such work can now be done on freshly collected material without culturing the microorganisms. This project will provide for (1) unambiguous identification of the dominant marine bacteria, in a way appropriate for comparing species compositions of both culturable and nonculturable populations, (2) selection of isolates that are most closely related to those dominating "real" marine populations, suitable for lab studies, (3) the development of molecular probes to define population distribu- tions in other locales and thus measure diversity within and between broadly different environments, (4) the possible use of these probes for single-cell hybridization studies for rapid microscopic identification of microorganisms in preserved samples from any marine environment.