Development of Ribosomal RNA-Based Oligonucleotide Probes for Green Ultraphytoplankton

基于核糖体 RNA 的绿色超浮游植物寡核苷酸探针的开发

• 批准号: 9202732
• 负责人: Marvin Fawley
• 金额: $ 15.8万
• 依托单位: North Dakota State University Fargo
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-15 至 1995-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9202732&HistoricalAwards=false
• 关键词: Development; Ribosomal; RNA; Based; Oligonucleotide

Microscopic, photosynthetic plankton are responsible for a significant portion on the primary productivity in low nutrient ocean waters. However, these ultraplankton which are present are poorly characterized. Algae cultured from these populations may not be representative of the actual dominant organisms. Attempts at describing the structure of deep-water communities by high performance liquid chromatography, flow cytometry, and electron microscopy have provided some important information, but suffer from quantitative and/or qualitative problems. This project plans to develop a different method to address the problem of identifying and quantifying photosynthetic ultraplankton by ribosomal RNA-based oligonucleotide probes. Short strands of oligonucleotides (radiolabelled or fluorescent) which hybridize to specific ribosomal-RNA sequences will be synthesized. Because of the abundance of rRNA in cells, these oligonucleotides can be used to probe mixed populations of whole cells. Probes specific for certain orders of green algae (especially the Mamiellales, which are known to be related to some small coccoid phytoplankton) will be produced and tested with a number of coccoid and flagellate algae. These probes should allow the enumeration of algae belonging to specific orders in field samples.

微小的光合作用浮游生物是低营养海水初级生产力的重要组成部分。然而，目前存在的这些超浮游生物的特征并不理想。从这些种群中培养出来的藻类可能不能代表实际的优势生物。用高效液相色谱、流式细胞仪和电子显微镜描述深水群落结构的尝试提供了一些重要的信息，但存在定量和/或定性的问题。该项目计划开发一种不同的方法来解决利用基于核糖体RNA的寡核苷酸探针来识别和量化光合作用超浮游植物的问题。将合成与特定核糖体-RNA序列杂交的短链寡核苷酸(放射性标记或荧光)。由于细胞中含有丰富的rRNA，这些寡核苷酸可以用来探测整个细胞的混合种群。将生产针对某些目的绿藻(特别是Mamiellales，已知与一些小型球状浮游植物有关)的探针，并用一些球状和鞭毛藻进行测试。这些探头应允许对野外样品中属于特定目的藻类进行计数。