ENVIRONMENTAL TOXICANTS AND NAD(P)H HOMEOSTASIS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 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. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.