Equipment for Automated Acquisition of DNA Sequence and DNA Fragment Size Data

美国国家科学基金会建议0140522--“DNA序列和DNA片段大小数据的自动获取设备”--范德比尔特大学的David E.McCauley博士获得了一笔赠款，用于支持购买自动DNA测序仪、DNA提取系统和支持计算机硬件。基地站从平板凝胶中收集DNA序列数据，并以序列分析软件可以解释的形式存储。该机器还可以收集和处理其他类型的DNA片段大小数据，如微卫星DNA遗传标记。在DNA序列和微卫星分析的情况下，聚合酶链式反应(PCR)的产物被荧光染料标记，并在电泳过程中按大小分离。该基准站的优势在于其96井的容量和自动加载允许高吞吐量，其配置使其同样适合收集序列和微卫星数据。相关的DNA提取系统将允许从大量样本中快速提取基因组DNA。这些机器收集的数据将被五名P.I.S(D.McCauley，J.Burke，D.Funk，C.Johnson和N.O.Pellmyr)用于进化和种群生物学的各种研究，这些研究依赖于大量的DNA序列和/或微卫星数据。将利用该设备产生的数据的一些具体研究项目包括以下内容。1)一项旨在利用DNA序列信息和多态微卫星标记对北美入侵植物Silene cangaris的种群遗传学进行研究的研究。比较北美和原产于欧洲的植物的遗传特征，应该表明入侵的机制或途径。2)几种丝兰植物及其授粉蛾的比较分子系统学研究。这项系统发育研究将有助于证明高度共同进化的植物传粉者系统是如何发展的。3)利用多态微卫星座位的研究，将有助于从野生近亲中定位与向日葵驯化相关的基因。4)基于DNA序列的褐斑潜蝇复合体适应辐射和寄主植物迁移研究。5)蓝藻模拟实验室种群中随着分子标记频率变化而产生的生物钟基因适应意义的研究。所有这些研究都将有助于理解基本的生物过程，如范围扩大、共同进化、寄主植物转移和生物钟的进化。然而，来自这些研究的知识也将有助于解决更多应用问题，如如何控制入侵物种，昆虫如何将食物从本地植物物种转移到具有经济价值的植物，如何提高驯化物种的基因操作效率，或者我们如何克服与生物节律紊乱相关的健康问题。最后，所有研究的结果将有助于生物信息学的新兴领域，因为每一项研究都将要求P.I.改进收集、存储和分析大量遗传信息的方法。

Abstract for NSF Proposal 0140522 - "Equipment for Automated Acquisition of DNA Sequence and DNA Fragment Size Data"A grant has been awarded to Dr. David E. McCauley at Vanderbilt University to support the purchase of an automated DNA sequencer, a DNA extraction system, and supporting computer hardware. The BaseStation gathers DNA sequence data from slab gels and stores it in a form that can be interpreted by sequence analysis software. The machine can also gather and process other types of DNA fragment size data such as microsatellite DNA genetic markers. In the case of both DNA sequence and microsatellite analysis, products of polymerase chain reaction (PCR) are labeled with a florescent dye and separated by size during electrophoresis. The advantage of the BaseStation is that its 96 well capacity and automated loading allows for high throughput, and its configuration makes it equally suited for gathering sequence and microsatellite data. The associated DNA extraction system will allow for rapid extraction of genomic DNA from a large number of samples. The data gathered by these machines will be used by five P.I.'s (D. McCauley, J. Burke, D. Funk, C. Johnson and N.O. Pellmyr) in a variety of studies in evolutionary and population biology which rely on large quantities of DNA sequence and/or microsatellite data. Some of the specific research projects that will make use of data generated by the equipment include the following. 1) A study designed to use both DNA sequence information and polymorphic microsatellite markers to investigate the population genetics of Silene vulgaris, a plant invasive in North America. Comparison of the genetic characteristics of the plant in North America and in its native Europe should indicate the mechanism or route of invasion. 2) A comparative molecular systematics study of several species of yucca plants and of the moths that pollinate them. This phylogenetic study will help document how highly coevolved plant-pollinator systems develop. 3) A study using polymorphic microsatellite loci that will help to map genes associated with the domestication of sunflowers from their wild relatives. 4) A DNA sequence based study of adaptive radiation and host plant shifts in the Neochlamisus bebbianae beetle complex. 5) A study of the adaptive significance of biological clock genes that follows changes in the frequency of molecular markers in model laboratory populations of cyanobacteria. All of these studies will contribute to understanding basic biological processes such as range expansions, coevolution, host plant shifts and the evolution of the biological clock. However, knowledge coming from these studies will also contribute to the solution of more applied problems such as how invasive species might be controlled, how insects shift feeding from native plant species to plants of economic value, how the genetic manipulation of domesticated species might be made more efficient, or how we might overcome health problems associated with biological rhythm disorders. Finally, the results of all of the studies will contribute to the emerging field of bioinformatics, in that each study will require the P.I. to refine methods of gathering, storing, and analyzing a large volume of genetic information.