Role of Sigma Facotrs in Heterologous Gene Expression

Sigma Facotrs 在异源基因表达中的作用

• 批准号: 6779247
• 负责人: ZAKEE L SABREE
• 金额: $ 2.87万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6779247
• 关键词: DNA directed RNA polymerase; Escherichia coli; R factors; Rhodospirillales; bacterial capsules; bacterial genetics; biotechnology; electroporation; functional /structural genomics; fusion gene; gel mobility shift assay; gene expression; genetic library; genetic manipulation; genetic mapping; genetic polymorphism; genetic promoter element; hybrid enzyme; microorganism interaction; nucleic acid sequence; polymerase chain reaction; predoctoral investigator; soil microbiology; transcription factor

DESCRIPTION (provided by applicant): Soil microbes are ubiquitous, diverse and impact global nutrient cycling and biogeochemical structuring, yet much of our current understanding is built largely on what is known about culturable ebacteria, which comprise < 1% of the total microbial diversity. Advances in culturing methods and culture-independent approaches are essential to gain a more complete understanding of the microbial world. Our laboratories have developed metagenomics, which is a culture-independent approach to identifying genes and activities from soil microbes, thereby broadening our understanding of the uncultured microbes. Functional metagenomics relies on heterologous expression of foreign genes in Escherichia coli and other host species, and therefore the role of transcriptional factors in heterologous expression is integral to metagenomic analysis. My overall hypothesis is that introduction and manipulation of sigma factors can expand the range of genes from cultured and uncultured bacteria expressed in E. coli. My sub-hypotheses to be tested are: 1) Acidobacterium capsulatum RpoD will increase expression of Acidobacterium genes in E. coli 2) Induction of E. coli heat shock sigma factor (RpoH) will increase the likelihood of identifying tellurite resistance genes from Rhodobacter sphaeroides and uncultured bacteria The goal of this research is to establish the influence of sigma factors on heterologous gene expression. My research will contribute to our understanding of sigma factors and gene regulation and resulting in a novel functional characterization of genes from cultured and uncultured bacteria.

描述（由申请人提供）：土壤微生物无处不在、多样化，影响全球养分循环和生物地球化学结构，但我们目前的大部分理解主要建立在对可培养细菌的了解上，可培养细菌占微生物总多样性的<1%。培养方法和独立于培养的方法的进步对于更全面地了解微生物世界至关重要。我们的实验室开发了宏基因组学，这是一种独立于培养物的方法，用于识别土壤微生物的基因和活性，从而拓宽我们对未培养微生物的理解。功能宏基因组学依赖于大肠杆菌和其他宿主物种中外源基因的异源表达，因此转录因子在异源表达中的作用是宏基因组分析不可或缺的一部分。我的总体假设是，引入和操纵西格玛因子可以扩大大肠杆菌中表达的培养和未培养细菌的基因范围。我要测试的子假设是： 1) 荚膜酸杆菌 RpoD 会增加大肠杆菌中酸杆菌基因的表达 2) 诱导大肠杆菌热休克因子 (RpoH) 将增加从球形红杆菌和未培养细菌中鉴定亚碲酸盐抗性基因的可能性 本研究的目的是确定西格玛因子对异源基因表达的影响。我的研究将有助于我们对西格玛因子和基因调控的理解，并对培养和未培养细菌的基因进行新的功能表征。