A WAVE Nucleic Acid Fragment Analysis System for Research and Education

丹佛大学的菲利普·丹尼尔森博士获得了一笔赠款，用于资助购买转基因核酸片段分析系统。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由美国国家科学基金会资助的发现新基因的研究，这些基因编码：(1)大脑中的内分泌激素；(2)细胞色素P450毒素代谢酶--这对控制农作物病虫害和携带疾病的生物体至关重要；3)可用于清理受重金属污染的有毒废物的微生物蛋白质；以及4)可用于识别和跟踪濒危物种遗传多样性的分子标记--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，识别突变还需要费力地筛选数百到数千个基因，以寻找DNA序列中的细微变化。用目前使用的直接测序法分析单个新基因，可能需要一天或更长时间才能完成。由这笔赠款资助的转基因波系统将把分析时间减少到2-4分钟/样本。发现和分析编码毒素分解相关蛋白的基因，以及与应激有关的神经肽，是几个研究项目的重点。由于这些基因经常以重复的形式存在，存在细微但关键的差异，因此将两个副本分离是至关重要的。WAVE系统将被用来通过对用于基因扩增反应的初始DNA池进行精确的大小分离来减少潜在的竞争非靶标基因片段的数量。该仪器的突变检测和片段捕获功能将用于提高识别和恢复这些相关基因的效率-即使在两个基因序列相差不到0.5%的情况下也是如此。在专注于保护生物学和微生物生态学的研究中，WAVE系统将提供一种极其灵敏的方法来分析物种间和物种内的DNA序列差异。最后，WAVE的高速基因分型能力将被用于从数百个样本中收集基因频率数据，用于大规模、多州保护遗传项目。除了丹佛大学的研究活动带来的好处外，还将在本科生和研究生阶段推进一系列以实验室和课堂为导向的教育目标。在以分子为导向的实验室课程中，好处将特别明显，这些课程是分子生物学理学士和文学士学位的核心。在更广泛的层面上，部门为高中生提供的生物技术课程和教师培训讲习班，通过提供最现代的基因分析方法之一的第一手经验，也将得到极大的加强，以促进中学一级的实践科学教育。鉴于这些项目的对象是城市和低收入学区的学生，而这些学生在自然科学领域的代表性一直较低，高中推广工作的好处将是不可估量的。简而言之，收购WAVE核酸片段分析系统将为高中、本科生和研究生的教育提供显著和直接的好处。

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.