AFFINITY ISOLATION & I-DIRT MS ANALYSIS OF E COLI O157:H7 SAKAI RNA POL COMPLEX

亲和隔离

• 批准号: 8169142
• 负责人: Brian T Chait
• 金额: $ 0.12万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169142
• 关键词: Affinity; Bacteria; Code; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA-Directed RNA Polymerase; Escherichia coli; Escherichia coli O157; Funding; Genes; Genome; Grant; Institution; Laboratories; Manuscripts; Mass Spectrum Analysis; Multienzyme Complexes; Proteins; Publishing; RNA; Research; Research Personnel; Resources; Source; Techniques; Transcription Initiation; Transcriptional Regulation; United States National Institutes of Health; Work; pathogenic Escherichia coli

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. Bacteria contain a single multi-subunit RNA polymerase that is responsible for the synthesis of all RNA. Previous studies on the Escherichia coli K-12 laboratory strain had identified a group of effector proteins that interact directly with RNA polymerase to modulate the efficiency of transcription initiation, elongation or termination. Here we have used a rapid affinity isolation technique to isolate RNA polymerase from the pathogenic Escherichia coli O157:H7 Sakai strain. We analysed the RNA polymerase enzyme complex using mass spectrometry and identified associated proteins. Although Escherichia coli O157:H7 Sakai contains more than 1,600 genes not present in the K-12 strain, many of which are predicted to be involved in transcription regulation, all of the identified proteins in this study were coded on the 'core' Escherichia coli genome. A Manuscript describing this work was published: Affinity isolation and I-DIRT mass spectrometric analysis of the Escherichia coli O157:H7 Sakai RNA polymerase complex. Lee DJ, Busby SJ, Westblade LF, Chait BT. J Bacteriol. 2008 Feb;190(4):1284-9.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 细菌含有单个多亚基RNA聚合酶，该聚合酶负责所有RNA的合成。先前对大肠杆菌K-12实验室菌株的研究已经确定了一组效应蛋白，该蛋白与RNA聚合酶直接相互作用以调节转录启动，伸长或终止的效率。在这里，我们使用了一种快速亲和力隔离技术，将RNA聚合酶与致病性大肠杆菌O157：H7 Sakai菌株分离。我们使用质谱法分析了RNA聚合酶复合物，并鉴定出相关的蛋白质。尽管大肠杆菌O157：H7 sakai包含K-12菌株中不存在的1,600多个基因，其中许多基因被预测与转录调节有关，但这项研究中所有鉴定的蛋白质均编码在“核心” Escherichia Coli基因组上。描述这项工作的手稿已出版： 大肠杆菌O157：H7 Sakai RNA聚合酶复合物的亲和力分离和I型乳光谱分析。 Lee DJ，Busby SJ，Westblade LF，Chait Bt。 J细菌。 2008年2月； 190（4）：1284-9。