Technology Core: Proteomics

我们将使用国家的最先进的，定量质谱为基础的蛋白质组学，以确定成员的蛋白质复合物在野生型和突变结核分枝杆菌菌株在各种生长和扰动条件下。关键蛋白复合物成员的系统和公正的定义，以及在各种生长和扰动条件下成员的变化，将提供关键的洞察到重要的基因的生物学功能的生长和生存的结核分枝杆菌。通过整合蛋白质组学、代谢组学和转录组学分析的结果，应该出现对生物体的生长和生存至关重要的相互作用和表达程序的全面画面。 我们还将使用最先进的，定量的全球蛋白质组学来分析结核分枝杆菌的营养缺陷型菌株，以了解编码蛋白质的基因和功能未知的非编码RNA在结核分枝杆菌中的作用。结核我们将选择生长变化发生之前的条件/时间点，以最大限度地减少蛋白质丰度变化是生长和存活变化的结果，而是直接归因于突变的可能性。将分析野生型和突变型细菌菌株 通过比较定量蛋白质组学分析。我们预计会发现几种不同类型的变化。我们将把变化与整个联盟的其他实验观察结果联系起来。

We will use state-of-the-art, quantitative mass spectrometry-based proteomics to define membership of protein complexes in wild-type and mutant Mtb strains under a variety of growth and perturbational conditions. Systematic and unbiased definifion of the membership of key protein complexes, and the changes in that membership under a variety of growth and perturbaional condtions, will provide critical insights into the biological function of genes important for growth and survival of Mtb. By integrating results of proteomics, metablomics and transcriptomic analyses, a comprehensive picture of the interactions and expression programs critical for growth and survival of the organism should emerge. We will also use state-of-the-art, quantitative global proteomics to profile auxotrophic strains of Mtb to understand the roles of genes encoding proteins and non-coding RNAs of unknown function in M. tuberculosis. We will choose condtions/time points just before changes in growth occur to minimize the possibility that changes in protein abundance are a consequence of changes in growth and survival but, instead, are directly attributable to the mutation. Both wild type and mutant bacteria strains will be analyzed by comparative quantitative proteomic analysis. We anticipate that we will find several different types of changes. We will correlate changes with observations from other experiments across the consortium.