A WAVE Nucleic Acid Fragment Analysis System for Research and Education

用于研究和教育的 WAVE 核酸片段分析系统

• 批准号: 0200484
• 负责人: Phillip Danielson
• 金额: $ 10.73万
• 依托单位: University of Denver
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0200484&HistoricalAwards=false
• 关键词: WAVE; Nucleic; Acid; Fragment; Analysis

A grant has been awarded to Dr. Phillip Danielson at the University of Denver to fund the purchase of a Transgenomic Nucleic Acid Fragment Analysis System. Based on established DNA size separation and mutation detection technology, this system will increase the quality and cost effectiveness of undergraduate/graduate research andeducation in molecular biology. Specific research programs that will benefit immediately include National Science Foundation-funded studies to identify novel genes that encode: (1) endocrine hormones in the brain; (2) cytochrome P450 toxin-metabolizing enzymes - which are critical to the control of crop pests and disease-carrying organisms; 3) microbial proteins that can be used to clean up of toxic waste sites contaminated with heavy metals and 4) molecular markers that can be used to identify and track genetic diversity in endangered species - work is conducted in collaboration with the Denver Zoological Gardens and Denver Museum of Science and Nature. Until recently, the identification of mutations required the laborious screening of hundreds to thousands of genes for subtle variations in DNA sequence. Analysis of a single novel gene by the direct sequence approach currently used, can require a day or more to complete. The Transgenomic WAVE System funded by this grant will reduce the analysis time to 2-4 minutes/sample. The discovery and analysis of genes that encode proteins involved in toxin breakdown, as well as neuropeptides linked to stress is the focus of several research programs. Since these genes often exist as duplicates with subtle but critical differences, it is essential that both copies be isolated. The WAVE system will be used to reduce the potential number of competing non-target gene fragments by precise size fractionation of the initial pool of DNA used for gene amplification reactions. The instrument's mutation detection and fragment capture functions will be used to increase the efficiency with which these related genes are identified and recovered - even where two genes sequences differ by less than 0.5%. In research focused on conservation biology and microbial ecology, the WAVE system will provide an extremely sensitive approach to the analysis of DNA sequence differences among and within species. Finally, the WAVE's high-speed genotyping capabilities will be used to gather gene frequency data from hundreds of samples for large-scale, multi-state conservation genetic projects. Beyond the benefit to the research activities at the University of Denver, a broad range oflaboratory and classroom-oriented educational goals will be advanced at both the undergraduate and graduate levels. Benefits will be particularly evident in the molecular-oriented laboratory courses that are at the heart of the Bachelor of Science and Bachelor of Arts degrees in Molecular Biology. On a broader level, department-sponsored biotechnology classes offered to high school students and teacher-training workshops that promote hands-on science education at the secondary school level will also be greatly enhanced by providing first-hand experience in one of the most modern methods of genetic analysis. The benefit to high school outreach efforts will immeasurable given that these programs target students in urban and low-income school districts who have traditionally been underrepresented in the natural sciences. In short, acquisition of theWAVE Nucleic Acid Fragment Analysis System will provide significant and immediatebenefits to education at the high school, undergraduate and graduate levels.

丹佛大学的菲利普·丹尼尔森博士获得了一笔赠款，用于购买一个转基因的 核酸片段分析系统。基于已建立的DNA大小分离和突变检测技术，该系统将提高分子生物学本科生/研究生研究和教育的质量和成本效益。将立即受益的具体研究计划包括国家科学基金会资助的研究，以确定编码以下内容的新基因：（1）大脑中的内分泌激素;（2）细胞色素P450毒素代谢酶-这对控制作物害虫和携带疾病的生物至关重要; 3）微生物蛋白质，可用于清理被重金属污染的有毒废物场地，4）分子标记，可用于识别和跟踪濒危物种的遗传多样性-这项工作是与丹佛动物园和丹佛科学与自然博物馆合作进行的。直到最近，突变的识别需要对数百到数千个基因进行艰苦的筛选，以寻找DNA序列中的细微变化。通过目前使用的直接测序方法分析单个新基因可能需要一天或更长时间才能完成。由该基金资助的转基因WAVE系统将把分析时间减少到2-4分钟/样本。发现和分析编码参与毒素分解的蛋白质以及与压力有关的神经肽的基因是几个研究项目的重点。由于这些基因通常以具有细微但关键差异的复制品存在，因此必须将两个副本分离。WAVE系统将用于通过对用于基因扩增反应的初始DNA池进行精确大小分级分离，减少竞争性非靶基因片段的潜在数量。该仪器的突变检测和片段捕获功能将用于提高识别和回收这些相关基因的效率-即使两个基因序列的差异小于0.5%。在保护生物学和微生物生态学研究中，WAVE系统将为分析物种之间和物种内部的DNA序列差异提供一种极其灵敏的方法。最后，WAVE的高速基因分型能力将用于从数百个样本中收集基因频率数据，用于大规模、多州的保护基因项目。除了有利于丹佛大学的研究活动之外，还将在本科和研究生阶段推进广泛的实验室和课堂教育目标。好处将是特别明显的分子为导向的实验室课程，是在科学和文学学士学位的分子生物学学士学位的核心。在更广泛的层面上，通过提供最现代的遗传分析方法之一的第一手经验，也将大大加强由系资助的高中生生物技术课程和促进中学科学教育实践的教师培训讲习班。考虑到这些项目针对的是城市和低收入学区的学生，这些学生传统上在自然科学方面的代表性不足，因此高中外展工作的好处是不可估量的。简而言之，WAVE核酸片段分析系统的收购将为高中、本科和研究生教育带来重大而直接的好处。