CORE B- Animal Models and Tissue Analysis

CORE B-动物模型和组织分析

• 批准号: 10268695
• 负责人: Dmitry Aleksandrovich Soloviev
• 金额: $ 36.07万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10268695
• 关键词: Address; Animal Model; Blood; Blood Vessels; Cell Adhesion; Cells; Complementary DNA; Elements; Equipment; Goals; Human Resources; IRAK1 gene; Individual; Integrins; Laboratories; Molecular; Mutagenesis; Physiological; Procedures; Protein Fragment; Proteins; Reagent; Signal Transduction; Structure-Activity Relationship; Talin; Training; animal tissue; experience; mutant; paxillin; programs; protein expression; protein purification

Project Summary The primary goal of this Program Project is to understand the molecular mechanisms by which the physiological and pathophysiological functions of integrins are regulated in blood and vascular cells. The “Protein Expression and Purification” Core B will assist each of the three projects realize this common goal by providing the proteins and cDNAs needed to analyze these molecular mechanisms. The proteins needed by the projects of the program are integrins, talin, kindlins, IRAK1, paxillin, and fragments of these proteins. The cDNAs needed are for expression of the integrins, talin, paxillin, kindlins, and IRAK1, mutant forms of and fragments of these proteins. By providing well-characterized proteins and cDNAs, this core creates reliable reagents that are useful to all three Projects of the Program. To address the structure-function relationships proposed in each project of the program requires substantial amounts of the wild-type and mutant forms of the target proteins. Both bacterial and mammalian expression of proteins is needed. The centralization of protein expression and mutagenesis avoids unneeded replication of procedures within individual laboratories, provides access to equipment that is needed for protein expression and purification, and allows these tasks to be performed efficiently and economically by experienced personnel. There is also a training element to Core B. The Core personnel will train fellows and technical staff within the individual projects to perform protein purification, bacterial and mammalian expression, mutagenesis strategies and in protein characterization approaches.

项目概要 该计划项目的主要目标是了解生理学的分子机制 整合素的病理生理功能在血液和血管细胞中受到调节。 “蛋白质表达 和纯化”核心 B 将通过提供蛋白质来协助这三个项目实现这一共同目标 以及分析这些分子机制所需的 cDNA。该计划项目所需的蛋白质 是整合素、talin、kindlins、IRAK1、paxillin 以及这些蛋白质的片段。所需的 cDNA 用于 整合素、talin、paxillin、kindlins 和 IRAK1、这些蛋白质的突变形式和片段的表达。 通过提供充分表征的蛋白质和 cDNA，该核心创建了对所有人都有用的可靠试剂 该计划的三个项目。解决每个项目中提出的结构-功能关系 程序需要大量的野生型和突变型的目标蛋白。两者都是细菌 并且需要哺乳动物表达蛋白质。蛋白质表达和诱变的集中化 避免在各个实验室内不必要地重复程序，提供对设备的访问 蛋白质表达和纯化所需的，并允许有效地执行这些任务 由经验丰富的人员在经济上进行。核心 B 也有一个培训要素。核心人员将接受培训 各个项目中的研究员和技术人员进行蛋白质纯化、细菌和哺乳动物 表达、诱变策略和蛋白质表征方法。