MRI/RUI-Acquisition of a Real-Time Quantitative PCR Instrument for Undergraduate Research and Teaching

在Vandergon、布鲁斯特、Nofziger Plank和Helm博士的监督下，佩珀代因大学获得了一笔赠款，用于购买研究仪器，以加强细胞和分子生物学的本科研究和教学计划。 由该基金资助的仪器旨在进行实时定量聚合酶链反应（QPCR）实验。 QPCR是一种技术，其中随着反应的进行，低拷贝数DNA或RNA样品被扩增并用荧光染料真实的实时检测。 实时检测扩增样品的能力在解决细胞和分子研究中的许多问题方面具有巨大价值，因为它允许进行准确的定量分析。 QPCR仪器将用于夏季本科研究项目和学术学期期间的荣誉研究项目。该仪器将促进在生物系目前的夏季计划的背景下进行的研究项目，部分通过NSF-REU网站赠款资助。 PI还将开发四个新的本科实验室教学模块，专门利用实时QPCR来解决样品中的低拷贝基因存在以及细胞，组织或生物体中的基因表达。传统上已经使用多种方法来测量正常基因表达和检测基因表达的变化，然而，量化传统的PCR和RT-PCR（逆转录PCR）测定是具有挑战性的，其易于出现模板转换错误和扩增错误。 QPCR技术的最新进展克服了这些问题，并允许在真实的时间实验中准确定量扩增的模板。 将使用QPCR仪器的具体研究项目包括：（1）两栖类光裂合酶基因表达对紫外线辐射的响应分析（Vandergon），（2）真核细胞中的应激反应信号和凋亡信号的检查（布鲁斯特），（3）哺乳动物细胞中Notch信号转导的表征（Nofziger Plank）和（4）在动物细胞培养模型系统中检查糖原合酶调节（Helm）。 此外，该仪器将用于将新概念引入核心细胞生物学和生物化学课程的实验室，几门高年级生物学选修课以及营养科学的相关课程。 与此相关的每个教师将开发一个新的本科实验室教学模块，探索样本中的基因存在，基因活性的时间变化或基因表达的诱导变化。 利用该仪器可实现四个实验教学模块：（1）大肠杆菌中间代谢基因反应分析;大肠杆菌（C.coli）中外阴发育相关基因的表达谱分析;（2）酵母菌中外阴发育相关基因的表达谱分析;（4）分析食物中是否含有转基因生物。 开发的实验室模块将在佩珀代因大学自然科学部的网站上免费提供。这笔赠款将促进佩珀代因大学细胞和分子生物学的本科生研究。 该工具将为学生自主研究提供一个强大的新工具。 该补助金将为本科生提供最新的分子和细胞生物学研究设备。 它还将帮助培养学生对细胞和分子生物学研究生学习的兴趣，并为有兴趣在生物技术行业就业的学生提供培训。 跨部门的连接将在生物化学计划，生物学计划和营养科学计划之间的研究和教学。 最终，这笔赠款将有助于培训科学家在尖端技术，以保持研究在佩珀代因大学在科学的前沿。

AbstractA grant has been awarded to Pepperdine University under the supervision of Drs. Vandergon, Brewster, Nofziger Plank, and Helm to acquire research instrumentation that will enhance the undergraduate research and teaching programs in cell and molecular biology. The instrument funded by this grant is designed to perform real-time quantitative polymerase chain reaction (QPCR) experiments. QPCR is a technique in which low copy number DNA or RNA samples are amplified and detected in real time with fluorescent dyes as the reactions are progressing. The ability to detect the amplified samples in real-time is of enormous value in addressing many questions in cell and molecular research because it allows for accurate quantitative analyses. The QPCR instrument will be utilized in both summer undergraduate research programs and in the Honor's research program during the academic semesters. This instrument will facilitate research projects carried out in the context of the biology department's current summer program funded in part through a NSF-REU site grant. The PI's will also develop four new undergraduate laboratory teaching modules that specifically utilize real-time QPCR in addressing low copy gene presence in samples, and gene expression in cells, tissues, or organisms. A variety of methods have been utilized traditionally for measurement of normal gene expression and detection of changes in gene expression, however, it is challenging to quantify traditional PCR and RT-PCR (reverse transcription PCR) assays which are prone to template conversion errors and amplification errors. Recent advances in QPCR technology overcome these problems and allow for accurate quantification of amplified templates in real time experiments. Specific research programs in which the QPCR instrument will be used are: (1) analysis of photolyase gene expression in response to UV radiation in amphibians (Vandergon), (2) examination of stress-response signals and apoptotic signals in eukaryotic cells (Brewster), (3) characterization of Notch signal transduction in mammalian cells (Nofziger Plank) and (4) examination of glycogen synthase regulation in animal cell culture model systems (Helm). In addition, this instrument will be used to introduce new concepts into the laboratories of the core cell biology and biochemistry courses, several upper-division biology elective courses, and related courses in nutritional science. Each faculty member associated with this grant will develop a new undergraduate laboratory teaching modules that explores gene presence within samples, temporal changes in gene activity, or induced changes in gene expression. The four laboratory teaching modules that will be implemented using this instrument include: (1) an analysis of intermediary metabolism gene responses of E. coli grown in different nutrient media, (2) an analysis of gene expression changes in response to environmental stress in yeast, (3) an analysis of the expression patterns of two genes involved in vulval development in C. elegans, and (4) an analysis of the presence of genetically modified organism (GMO) material in foods. The developed laboratory modules will be made available freely on the Natural Science Division Web site at Pepperdine University.This grant will foster undergraduate research in cell and molecular biology at Pepperdine University. This instrument will provide a powerful new tool for use in student-generated independent research. The grant will provide undergraduate students with access to the latest equipment for research in molecular and cellular biology. It will aid also in generating student interest in graduate studies in cell and molecular biology and will provide training to students interested in employment in the biotechnology industry. Cross-department connections will be made in research and teaching between the biochemistry program, biology program, and nutritional sciences program. Ultimately, this grant will assist in training scientists in cutting edge techniques to keep research at Pepperdine University at the forefront of science.