Molecular Resources Core

总结： 核心C，即分子资源核心，将为所有四个项目提供分子试剂和服务，对完成计划项目中每个提案的具体目标至关重要。分子资源核心的提议是原始核心C的延续。核心C的总体功能是 提供分子试剂和专业知识，包括产生缺失和定点突变体并掺入所需的表达载体，产生用于转导原代内皮细胞的腺病毒和逆转录病毒构建体，敲除小鼠的基因分型，设计和产生适当的基于siRNA的构建体，对从其他研究者获得的构建体进行测序和验证，对项目人员进行分子生物学培训，以及监督和维护用于有效转染内皮细胞的仪器。核心C还将提供蛋白质组学分析和氧化剂测量的专业知识。这 核心将由兰德尔·A博士指导。Skidgel和分子试剂和服务将由高级研究专家（Tiffany Sharma博士）、研究专家（Debra Salvi）和研究助理教授（Fulong Tan博士）提供。Neil Kelleher博士，蛋白质组学卓越中心教授兼主任， 将建议，监督和进行蛋白质组学分析的质谱。Marcelo Bonini博士，助理教授，将提供专业知识，建议，并将进行过氧亚硝酸盐和其他氧化剂的测量，以剖析它们在调节肺血管内皮屏障功能中的作用。巩固这些努力， 核心C将提高完成每个项目中概述的研究任务的速度和效率，并统一方法和方法，以便可以比较每个项目的结果。此外，它将允许严格的质量控制和分子试剂的一致性，以供各种 项目此外，集中式核心C的一个额外的重要好处是，与每个项目都产生自己的分子资源的情况相比，它将显着减少费用。核心领导人与高级研究专家和项目领导人的定期会议将确保协调 以及分子试剂生成的优先级，并确保所需试剂及时提供给组件项目。

Summary: Core C, the Molecular Resources Core, will provide molecular reagents and services to all four projects and will be crucial to the completion of the specific aims of each proposal in the Program Project. The proposal for a Molecular Resources Core is a continuation of the original Core C. The overall function of Core C is to provide molecular reagents and expertise, including generation of deletion and site-directed mutants and incorporation into the desired expression vectors, generation of adenoviral and retroviral constructs for transducfion of primary endothelial cells, genotyping of knockout mice, designing and generating appropriate siRNA based constructs, sequencing and verification of constructs obtained from other investigators, molecular biology training to project personnel, and overseeing and maintaining instrumentation for efficient transfection of endothelial cells. Core C will also provide expertise for proteomic analysis and oxidant measurements. This core will be directed by Dr. Randal A. Skidgel and molecular reagents and services will be provided by a Senior Research Specialist (Dr. Tiffany Sharma), Research Specialist (Debra Salvi) and Research Assistant Professor (Dr. Fulong Tan). Dr. Neil Kelleher, Professor and Director of the Proteomics Center of Excellence, will advise, oversee and carry out proteomic analysis by mass spectrometry. Dr. Marcelo Bonini, Assistant Professor, will provide expertise, advice and will carry out measurement of peroxynitrite and other oxidants to dissect their roles in regulating lung vascular endothelial barrier barrier function. Consolidating these efforts in Core C will result in increased speed and efficiency in accomplishing the research tasks outlined in each of the projects and uniformity of approaches and methods so that results from each project can be compared. In addition, it will allow strict quality control and consistency of molecular reagents to be used by the various projects. Furthermore, an added and important benefit of the centralized Core C will be to markedly decrease expenses compared to a scenario in which each project would generate its own molecular resources. Regular meetings of the Core Leader with the Senior Research Specialist and project leaders will assure coordination and prioritization of the generation of molecular reagents and assure that the desired reagents are provided in a timely manner to the component projects.