EQUIPMENT-HPLC

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。 在项目2中，我们研究了宿主磷脂酰胆碱的细菌分解代谢与宿主定植相关细菌基因的诱导之间的联系。这个试点项目旨在购买设备来跟踪磷脂的降解并监测PC分解代谢途径中中间产物的积累。虽然我们最初寻求购买高效液相色谱仪，但在与Aligent和岛津的技术专家多次互动后，很明显LC-MS是该项目所需的敏感和定量研究的更合适的技术，我们的实验室没有能力投入资金或时间来建立LC-MS能力，因为Cobre研究人员已经可以使用这项技术。有鉴于此，我们成功地试行了一些比色和荧光分析方法来检测这些化合物。为了使我们的实验室能够在实验室的常规基础上使用这些分析，我们使用试点资金购买了一台微型滴定板读取器，该读取器具有必要的软件和功能，使我们能够监测PC分解代谢和分解代谢中间体。我们的项目还将重点放在参与细菌硝酸盐呼吸的基因以及它们与宿主定植的联系上。该分光光度计还将用于通过比色分析跟踪氮氧化物。

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. In Project 2, we examine the connection between the bacterial catabolism of host phosphatidylcholine and the induction of bacterial genes involved in host colonization.This pilot project was aimed at purchasing equipment to follow the degradation of phospholipids and to monitor the accumulation of intermediates in the PC catabolic pathway. While we initially pursued the purchase of an HPLC, after multiple interactions with technical experts from Aligent and Shimadzu, it became clear that LC-MS was the more appropriate technique for the sensitive and quantitative studies that would be required by this project and our lab is not equipped to invest the funds or time to establish LC-MS capabilities as this technology is already available to COBRE investigators. In light of this, we successfully piloted a number of colorimetric and fluorescent assays for the detection of these compounds. To enable our lab to use these assays on a routine basis in our laboratory, we have used pilot funds to purchase a microtiter plate reader with the necessary software and capabilities that will enable us to monitor PC catabolism and catabolic intermediates. Our project has also turned to focus on genes involved in bacterial nitrate respiration and their connection to host colonization. The spectrophotometer will also be used to follow nitrogen oxides via colorimetric assays.