GENE EXPRESSION IN BORRELIA BURGDORFERI

伯氏疏螺旋体的基因表达

• 批准号: 7958570
• 负责人: RAMESH RAMAMOORTHY
• 金额: $ 5.8万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7958570
• 关键词: Biological Assay; Borrelia; Borrelia burgdorferi; Characteristics; Computer Retrieval of Information on Scientific Projects Database; Core Protein; Cytosine; Deletion Mutation; Discrimination; Elements; Funding; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Grant; Green Fluorescent Proteins; Guanosine; Infection; Institution; Methods; Organism; Primates; Reporter; Research; Research Personnel; Resources; Site; Source; Tissue-Specific Gene Expression; United States National Institutes of Health; Variant; base; promoter

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. Background & Aims: Regulation of gene expression in the Lyme disease spirochetes is of fundamental importance to establishing a successful infection. In this study, we investigated the functional elements of the flaB promoter of Borrelia burgdorferi. Our goal was to understand the basis for promoter discrimination in this organism. Promoters constitute one of the basic determinants of differential gene expression in this organism. Methods: Promoter function was examined in a high-passage variant of strain JD1 using a set of 5' deletions and mutations within the flaB promoter. Expression from the modified flaB promoters was assayed using the gene for green fluorescent protein (gfp) as a reporter. Results: Although the -35 element of the promoter stimulated promoter activity, its disruption did not negate expression. Sequence upstream of the -35 had no effect on expression. The -35/-10 spacer region composed of a T-rich sequence was critical for optimal promoter function. Surprisingly, a Cytosine at the -13 site was found to be more favorable for transcription compared to a Guanosine at the same site. Conclusions: Based on these results and other characteristics, we propose that the B. burgdorferi flaB promoter is an example of an extended -10 promoter. Further, the T-rich spacer is a key element of the flaB promoter that contributes to the abundance of the flagellar core protein in Borrelia species.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 背景和目的：莱姆病螺旋体基因表达的调节对于建立成功的感染至关重要。在本研究中，我们研究了伯氏疏螺旋体flaB启动子的功能元件。我们的目标是了解该生物体中启动子歧视的基础。启动子构成该生物体差异基因表达的基本决定因素之一。方法：使用 flaB 启动子内的一组 5' 缺失和突变在菌株 JD1 的高传代变体中检查启动子功能。使用绿色荧光蛋白（gfp）基因作为报告基因来分析修饰的flaB启动子的表达。结果：虽然启动子的-35元件刺激了启动子活性，但其破坏并没有消除表达。 -35上游的序列对表达没有影响。由富含 T 的序列组成的 -35/-10 间隔区对于最佳启动子功能至关重要。令人惊讶的是，与同一位点的鸟苷相比，-13 位点的胞嘧啶被发现更有利于转录。结论：基于这些结果和其他特征，我们提出伯氏疏螺旋体 flaB 启动子是延伸的 -10 启动子的一个例子。此外，富含 T 的间隔区是 flaB 启动子的关键元件，有助于伯氏疏螺旋体物种中鞭毛核心蛋白的丰度。