Protein Engineering and Expression Core

C.核心单元描述： C.1.理论基础和目标 参与该计划项目的三个实验室中的每一个都将表达从血清蛋白(ApoA1)和酶(PON1、LCAT和MPO)到细胞质核糖体结构蛋白(如L13a)、酶(eNOS、nNOS和Akt)及其子集的突变形式的不同类别的蛋白质，以便执行项目中描述的后续酶分析，并利用生物物理和计算化学核心(核心C)中详细说明的各种复杂仪器进行分析。这个 蛋白质工程和表达核心的主要目标是为大量的蛋白质表达提供一种集中的手段，从而促进对每个项目感兴趣的蛋白质进行相对复杂和多样化的生物物理研究。核心还将作为一个教育设施，培训计划项目实验室的成员如何使用各种策略优化蛋白质表达，以及产生蛋白质表达所需的特定等位基因的突变。核心还将提供蛋白质纯化方面的培训。集中这些方面的工作和培训预计将极大地提高参与每个项目的科学家的效率和生产力。

C. Description of Core Unit: C.1. Rationale and Objectives Each of the three laboratories involved in the Program Project will express distinct classes of proteins from serum proteins (ApoA1) and enzymes (PON1, LCAT, and MPO) to cytoplasmic ribosomal structural proteins (like L13a), enzymes (eNOS, nNOS, and Akt) and mutant forms of a subset of these in order to perform subsequent enzymatic assays as described in the Projects and analysis utilizing diverse and sophisticated instrumentation detailed in the Biophysical and Computational Chemistry Core (Core C). The principal objective of the Protein Engineering and Expression Core is to provide a centralized means for high volume protein expression thereby facilitating the relatively sophisticated and diverse biophysical studies on the proteins of interest for each Project The Core will also function as an educational facility for training members of the Program Project laboratories on ways to optimize protein expression using various strategies and also in the generation of mutants of specific alleles desired for protein expression. Training in the purification of proteins will also be provided by the Core. Centralizing these aspects of the work and training is expected to greatly enhance efficiency and productivity of the scientists participating in each project.