RUI: Acquisition of DNA Sequencer, Synthesizer and Spectrophotometer

RUI：购置 DNA 测序仪、合成仪和分光光度计

• 批准号: 0116617
• 负责人: David Lawlor
• 金额: $ 11.59万
• 依托单位: Rochester Institute of Tech
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2005-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0116617&HistoricalAwards=false
• 关键词: RUI; Acquisition; DNA; Sequencer; Synthesizer

Abstract A grant has been awarded to Dr. Lawlor at Rochester Institute of Technology to purchase three pieces of research equipment: a Genetic Analyzer, a DNA synthesizer and a diode array spectrophotometer. These complementary pieces would form an equipment suite that would facilitate the research programs of four investigators in the departments of biological sciences and chemistry. The research areas are diverse; they range from molecular characterization of immune response molecules in the great apes to creation of bacterial-resistant transgenic plants. They also include the functional characterization of bacterial enzyme systems, as well as the definition of closely related Central-American iguana species. The immune response molecules regulate the activity of natural killer cells, a lymphoid cell population instrumental in detecting virus-infected cells. The transgenic plant project explores the possibility of creating plants with increased production of a naturally occurring molecule that protects against bacterial parasites. The function of threonine dehydrogenase, an essential enzyme in a key metabolic pathway for many bacterial species, will be better understood after molecular characterization of its variant forms. The iguana project is intent on enumerating the species central Mexico and determining how similar they are to one another. The projects, although disparate, share the feature that they are basic biological research projects that would benefit by being able to characterize and sequence the relevant genes. The suite of equipment will permit the requisite DNA sequencing experiments so that the apes can be tested for the presence of the natural killer genes. If present, precise comparisons can be made against those found in human. The plant transgene, created using recombinant DNA techniques, needs to be sequenced to insure proper construction of the gene. Without this quality control assessment, subsequent experiments cannot be undertaken. Preliminary sequence information obtained for gram-negative bacteria, other than E. coli, reveals major differences in the nucleotide sequence for threonine dehydrogenase. These differences will be completely defined by sequencing the gene from the various bacterial species. Earlier field trips to Mexico provided the samples required for DNA sequence analysis of genes, mitochondrial and nuclear, that will form the database for estimating relatedness amongst the species. Although the DNA sequencer is the central equipment item, the DNA synthesizer will allow oligonucleotide production for the sequencing experiments and the spectrophotometer is necessary for accurate estimations of reagent amounts. The scientific contributions from the four projects will be enhanced due to the expanded capability provided by the equipment. Experiments that couldn't be undertaken previously can now be performed in the respective laboratories. It is expected that a better definition of the evolution of the natural killer genes in humans and apes will be one result. Also, testing the hypothesis that transgenic plants are superior to non-transgenic plants in regards to bacterial invasion will be possible. The characterization of the threonine dehydrogenase gene will provide insights as to the regions of the molecule critical for its function. Finally, an understanding of the biogeography of mesoamerica will be increased from the description of the iguana species that are indigenous to that area.

摘要 罗彻斯特理工学院的 Lawlor 博士获得一笔资助，购买了三件研究设备：遗传分析仪、DNA 合成仪和二极管阵列分光光度计。 这些互补的部件将形成一个设备套件，促进生物科学和化学系四名研究人员的研究项目。 研究领域多元化；它们的范围从类人猿免疫反应分子的分子特征到抗细菌转基因植物的创建。 它们还包括细菌酶系统的功能特征，以及密切相关的中美洲鬣蜥物种的定义。 免疫反应分子调节自然杀伤细胞的活性，自然杀伤细胞是一种有助于检测病毒感染细胞的淋巴细胞群。 转基因植物项目探索了创造植物的可能性，以增加天然存在的分子的产量，以防止细菌寄生虫。 苏氨酸脱氢酶是许多细菌物种关键代谢途径中的必需酶，在对其变体形式进行分子表征后，我们将更好地了解苏氨酸脱氢酶的功能。 鬣蜥项目旨在列举墨西哥中部的物种并确定它们之间的相似程度。 这些项目虽然不同，但都有一个共同的特点：它们都是基础生物学研究项目，能够通过对相关基因进行表征和测序而受益。这套设备将允许进行必要的DNA测序实验，以便可以测试猿类中是否存在自然杀伤基因。 如果存在的话，可以与人类中发现的进行精确比较。 使用重组 DNA 技术创建的植物转基因需要进行测序，以确保基因的正确构建。 如果没有这种质量控制评估，后续实验就无法进行。 除大肠杆菌外，获得的革兰氏阴性细菌的初步序列信息揭示了苏氨酸脱氢酶的核苷酸序列的主要差异。 这些差异将通过对不同细菌物种的基因进行测序来完全确定。 早些时候对墨西哥的实地考察提供了基因、线粒体和核 DNA 序列分析所需的样本，这些样本将形成用于估计物种之间相关性的数据库。 尽管DNA测序仪是核心设备，但DNA合成仪将允许生产用于测序实验的寡核苷酸，并且分光光度计对于准确估计试剂量是必需的。由于设备提供的能力扩展，这四个项目的科学贡献将得到增强。 以前无法进行的实验现在可以在各自的实验室进行。 预计结果之一将是对人类和猿类自然杀伤基因进化的更好定义。 此外，检验转基因植物在细菌入侵方面优于非转基因植物的假设也是可能的。 苏氨酸脱氢酶基因的表征将提供对其功能至关重要的分子区域的见解。 最后，通过对该地区本土鬣蜥物种的描述，将加深对中美洲生物地理学的了解。