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Transcriptional Regulation of Secondary Metabolism

次级代谢的转录调控

基本信息

批准号：
7448499
负责人：
MARY A O'CONNELL
金额：
$ 24.61万
依托单位：
NEW MEXICO STATE UNIVERSITY LAS CRUCES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7448499
关键词：
3&apos Untranslated Regions Acrylamide Acrylamides Adsorption Affinity Chromatography Aliquot Alleles Amino Acid Sequence Amino Acid Sequence Databases Anabolism Anthocyanins Antibodies Antigens Applied Research BHLH Protein Binding Binding Sites Biochemical Genetics Bioinformatics Biological Assay Capsaicinoid Capsicum Cell Nucleus Cells Chalcone synthase Chemicals Chile Chimeric Proteins Class Cloning Code Complementary DNA Complex Computer Analysis Computer software Condition Consensus Sequence Conserved Sequence Control Locus DNA DNA Binding Domain DNA Restriction Enzymes DNA Sequence Detection Development Electron Microscope Electrons Electrophoresis Electrophoretic Mobility Shift Assay Elements Engineering Enzymes Epitopes Euchromatin Event Experimental Designs Family member Figs - dietary Flowers Formaldehyde Fruit Gel Genbank Gene Expression Gene Expression Regulation Gene Family Gene Targeting Genes Genetic Transcription Genome Genome Mappings Genomics Genotype Goals Gold Colloid Hand Heterochromatin Human Resources Image Immune Sera Immunohistochemistry In Vitro Incubated Indium Individual Introns Jupiter Label Laser Scanning Confocal Microscopy Lasers Left Length Libraries Localized Location MYB gene Manufacturer Name Maps Measurable Measures Membrane Metabolic Metabolism Methods Microscope Microscopy Modeling Molecular Weight Nature Nested Genes Nested PCR New Mexico Northern Blotting Nuclear Nuclear Protein Nuclear Proteins Numbers Nylons Oils Online Systems Open Reading Frames Organ Oryctolagus cuniculus Outcome Pathway interactions Pattern Peptide Sequence Determination Peptides Pharmacologic Substance Physiological Placenta Placental Extracts Plant Leaves Plant Proteins Plant Roots Plants Plasmids Polymerase Chain Reaction Population Positioning Attribute Post-Translational Protein Processing Potato Preparation Procedures Progress Reports Promega Promoter Regions Protein Binding Protein Overexpression Proteins Protocols documentation Publishing Quantitative Trait Loci RNA Reaction Reading Recombinant Proteins Recombinants Regulation Regulator Genes Regulatory Element Relative (related person)Reporting Resolution Sampling Scanning Scheme Score Screening procedure Seeds Sepharose Sequence Alignment Sequence Analysis Series Serum Shapes Signal Transduction Site Solanaceae Solutions Southern Blotting Specificity Specimen Staging Standards of Weights and Measures Structural Genes Structure System Testing Tissues Tobacco Tomatoes Transcript Transcription factor genes Transcriptional Regulation Translating Translations Transmission Electron Microscopy Untranslated Regions Variant Walking Western Blotting Work base cDNA Library cell type chromatin immunoprecipitation crosslink design desire in vivo interest light microscopy link protein member myb Genes particle polyclonal antibody promoter research study sample fixation size stem synthetic enzyme tool trait transcription factor transmission process

项目摘要

Capsaicinoids are products of the phenylpropanoid pathway, a central and ubiquitous pathway in plant secondary metabolism. The products of this pathway are diverse, perform a multitude of functions in the plant, and are the chemical basis for numerous pharmaceutical agents. There are heritable differences in the amount and type of capsaicinoids that accumulate in fruit of different Capsicum lines; these differences are controlled by other genes, whose nature is also unknown. Our long-term goal is to understand the genetic, biochemical, physiological and environmental influences on the biosynthesis of capsaicinoids. We have demonstrated that there is differential transcription of the biosynthetic genes for capsaicinoids in the placenta of Capsicum spp. The pattern of transcript accumulation suggests that there are transcription factors that will bind to common cis elements in the promoters of these genes. None of the structural genes that we have characterized appear to be associated with the loci that control pungency, either the C locus or the QTL locus, caps. We anticipated that the genes that control pungency are likely regulatory, and now we are in a position to formally test that hypothesis. This SCORE competing renewal proposal outlines a series of experiments to isolate and characterize candidate transcription factors. We propose to characterize the cis-elements of the structural genes as well as candidate regulatory genes for the capsaicinoid pathway. The specific aims of this proposal are: 1) Complete a detailed characterization of the promoter regions of the structural genes on the capsaicinoid biosynthetic pathway; 2) Isolate and characterize genes for transcription factors expressed uniquely in chile placenta; 3) Determine the cell specific pattern of expression for these transcription factors using immunolocalization; 4) Determine if these transcription factors regulate/bind to promoters of capsaicionoid biosynthetic genes. To meet these objectives we will isolate placental specific transcription factors from habanero using a combination of cDNA library screening and PCR amplification. We have a bZIP candidate in hand and expect to isolate representative members of the myb and bHLH classes. Those trans-factors that are localized for accumulation in the nucleus of placental epidermal cells will be used in in vitro (gel shift) and in vivo (chromatin immunoprecipitation) assays to demonstrate binding.

辣椒素类化合物是苯丙素类化合物代谢途径的产物，苯丙素类化合物代谢途径是植物次生代谢的中心和普遍存在的途径。该途径的产物是多样的，在植物中执行多种功能，并且是许多药剂的化学基础。在不同辣椒品系果实中积累的辣椒素的数量和类型存在遗传差异;这些差异由其他基因控制，其性质也是未知的。我们的长期目标是了解辣椒素生物合成的遗传，生化，生理和环境影响。我们已经证明辣椒素生物合成基因在辣椒属植物胎盘中存在差异转录。转录物积累的模式表明，有转录因子，将结合到共同的顺式元件在这些基因的启动子。我们所描述的结构基因中没有一个似乎与控制辛辣的基因座（C基因座或QTL基因座，caps）相关。我们预计控制辛辣的基因可能是调节性的，现在我们可以正式验证这一假设。这个SCORE竞争更新提案概述了一系列分离和表征候选转录因子的实验。我们建议描述结构基因的顺式元件以及辣椒素类物质途径的候选调控基因。本研究的具体目标是：1）完成辣椒素生物合成途径结构基因启动子区的详细鉴定; 2）分离和鉴定在智利胎盘中独特表达的转录因子基因; 3）使用免疫定位确定这些转录因子的细胞特异性表达模式; 4）确定这些转录因子是否调节/结合辣椒素生物合成基因的启动子。为了满足这些目标，我们将使用cDNA文库筛选和PCR扩增的组合从哈巴内罗分离胎盘特异性转录因子。我们手头有一个bZIP候选者，并期望分离出myb和bHLH类的代表性成员。那些定位于胎盘表皮细胞核中积累的反式因子将用于体外（凝胶移位）和体内（染色质免疫沉淀）测定，显示绑定。