IN VIVO STUDY OF TRANSCRIPTIONAL REGULATION IN BACILLI BY FCS

FCS对杆菌转录调控的体内研究

• 批准号: 8171006
• 负责人: CATHERINE A ROYER
• 金额: $ 0.47万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171006
• 关键词: Arts; Bacillus (bacterium); Bacillus subtilis; Carbon; Chimeric Proteins; Computer Retrieval of Information on Scientific Projects Database; Engineering; Enzymes; Fluorescence; Funding; Genome; Gram-Positive Bacteria; Grant; Image; In Vitro; Institution; Investigation; Metabolic; Production; Research; Research Personnel; Resources; Scanning; Source; Spectrum Analysis; Techniques; Transcription Repressor/Corepressor; Transcriptional Regulation; United States National Institutes of Health; Work; in vivo; promoter; single molecule; two-photon

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. This project proposes the study of transcriptional regulation in Bacillus subtilis by direct, in vivo, observation using state-of-the-art two-photon fluorescence correlation spectroscopy. The project involves the study of two transcriptional repressors from B. subtilis, CggR and CcpN. Much work has been done in vitro on CggR and CcpN (Zorrilla et al, 2007a; Zorrilla et al, 2007b; Zorrilla et al, 2008a; Zorrilla et al, 2008b) since their discovery in the sequencing of the B. subtilis genome(Kunst et al, 1997). Both transcriptional repressors control opposite directions in the carbon metabolic cycle in gram positive bacteria through the production of metabolic enzymes. We propose to directly observe transcriptional regulation in vivo at the single molecule level. Using genetically engineered strains of B. subtilis, containing fluorescent protein fusions with CggR, CcpN and related factors under native promoters, we will apply the techniques of point and scanning two photon fluorescence correlation spectroscopy (FCS and sFCS), Number and Brightness (N&B) and raster image correlation spectroscopy (RICS). We will elucidate mechanisms of transcriptional regulation that cannot be observed by in vitro investigation.

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。 该项目提出了使用最先进的两光子荧光相关光谱的直接观察到枯草芽孢杆菌对枯草芽孢杆菌中转录调节的研究。 该项目涉及两个转录的研究 来自枯草芽孢杆菌，CGGR和CCPN的阻遏物。 自从他们在B. uttilis基因组的测序中发现以来，在CGGR和CCPN上已经在CGGR和CCPN上完成了很多工作（Zorrilla等，2007a; Zorrilla等，2007b; Zorrilla等，2008a; Zorrilla et al，2008b）。 两种转录阻遏物通过代谢酶的产生在革兰氏阴性细菌中的碳代谢周期中控制相反的方向。 我们建议在单分子水平上直接观察体内转录调节。 使用枯草芽孢杆菌的基因工程菌株，在天然启动子下含有荧光蛋白融合及其在天然启动子下的荧光蛋白融合，我们将应用点技术并扫描两个光子荧光光谱光谱光谱（FCS和SFCS），数量和亮度（N＆B）和Raster ImagerS Imager Corelation Corelation Spectrapsspept（Raster Image Corelation Spectrations（raster）。 我们将阐明无法通过体外研究观察到的转录调节机制。