ENVIRONMENTAL TOXICANTS AND NAD(P)H HOMEOSTASIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 INBRE梅维尔州立大学研究项目的长期目标是为本科生提供具有挑战性和回报的生物医学研究经验。通过Brin第一阶段资金，我们已经开始纳入创新的荧光技术，这是由当地一家高科技企业在NSF和NIH的资助下开发的。这项技术在产生高质量的荧光寿命信息方面取得了突破。将研究其在等位基因特异性SNP基因分型和实时聚合酶链式反应衰变中的应用。我们的假设是，荧光寿命方法将允许多重PCR和SNP检测，与目前商业上可用的设备和试剂盒相比，可以在更多的目标上以更低的成本、更高的灵敏度和选择性同时进行检测。我们的研究计划有四个具体目标： 1)表征环境对候选荧光团(自由形式和与寡核苷酸结合)的荧光性质的影响，这些候选荧光团是基于寿命的RT-PCR探针的候选者。 2)建立了利用寿命荧光探针监测单一和多重RT-PCR分析的方法学。 3)建立利用寿命荧光探针进行SNP检测的方法学。 4)通过与迪金森州立大学的INBRE小组、海龟山区社区学院和南大生物学核心设施的合作，将LIFE探针方法应用于真实世界样本的RT-PCR和SNP基因分型。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The long-term goal of the INBRE Mayville State University research program is to offer undergraduate students a challenging and rewarding experience directed toward biomedical research. Through BRIN Phase I funding, we have begun to incorporate innovative fluorescence technology, which has been developed by a local high tech business with funding from the NSF and NIH. This technology provides a breakthrough in the generation of high quality fluorescence lifetime information. Applications in allele-specific SNP genotyping and real-time PCR decay will be investigated. Our hypothesis is that the fluorescence lifetime methodology will allow multiplexed PCR and SNP detection that can be simultaneously performed on a greater number of targets with less expense, higher sensitivity and selectivity than is provided by equipment and kits that are currently available commercially. Our research plan has four specific aims: 1) characterize the environmental influence on the fluorescent properties of candidate fluorophores (both in their free form and bound to an oligonucleotide) that are candidates for lifetime-based RT-PCR probes. 2) develop methodology using the lifetime fluorescent probes to monitor single and multiplex RT-PCR analysis. 3) develop methodology using the lifetime fluorescent probes to perform SNP detection. 4) Apply the lifetime probe methodology to RT-PCR and SNP genotyping on real-world samples through collaboration with the INBRE groups at Dickinson State University, Turtle Mountain Community College, and the Biology Core Facility at NDSU.