Single-Cell Gel/Comet Assay Applied to the Analysis of Ultraviolet Induced DNA Damage and Kinetics of Repair in Phytoplankton

单细胞凝胶/彗星测定应用于浮游植物紫外线诱导 DNA 损伤和修复动力学的分析

• 批准号: 9712901
• 负责人: Miguel Sastre
• 金额: $ 5万
• 依托单位: University of Puerto Rico at Humacao
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-15 至 1998-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9712901&HistoricalAwards=false
• 关键词: Single; Cell; Gel; Comet; Assay

The stratospheric ozone layer has been diminished as a consequence of anthropogenic release of chlorofluorocarbons. The thinning ozone results in increased levels of harmful ultraviolet B (UV-B) radiation relative to less harmful ultraviolet A (UV-A) and visible light (VIS). Ultraviolet damage to marine phytoplankton cells have been limited to bulk calculations which do not take into consideration variability of damage between individual cells. The single cell gel (SSG) comet assay is capable of measuring DNA damage and repair in individual cells. We propose to use this technique to measure the effects of UV-B radiation on DNA damage in marine phytoplankton species and in their capacity to repair. Phytoplankton growing under culture conditions will be exposed in the laboratory to ambient irradiance levels of UV-B, UV-A and VIS to stimulate present and enhanced UV radiation. The ability of phytoplankton cells to repair DNA will be estimated to subsequent exposure to white light, to induce repair; and yellow light, to suppress repair. Results of these experiments will make an important contribution regarding the usefulness of a novel method to measure UV-induced DNA damage and repair in phytoplankton.

由于氯氟烃的人为排放，平流层臭氧层已经减弱。臭氧变薄导致有害的紫外线B(UV-B)辐射水平相对于危害较小的紫外线A(UV-A)和可见光(VIS)增加。紫外线对海洋浮游植物细胞的损害仅限于批量计算，没有考虑到单个细胞之间损害的可变性。单细胞凝胶(SSG)彗星试验能够测量单个细胞的DNA损伤和修复。我们建议使用这项技术来测量UV-B辐射对海洋浮游植物物种DNA损伤及其修复能力的影响。在培养条件下生长的浮游植物将在实验室暴露在UV-B、UV-A和VIS的环境辐射水平下，以刺激当前和增强的紫外线辐射。浮游植物细胞修复DNA的能力将被估计为随后暴露在白光下，诱导修复；以及黄光，抑制修复。这些实验的结果将对测量UV诱导的浮游植物DNA损伤和修复的新方法的有用性做出重要贡献。