REGULATION AND EVOLUTION OF MULTIGENE FAMILIES

多基因家族的调控和进化

• 批准号: 3272129
• 负责人: FOTIS C KAFATOS
• 金额: $ 23.19万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-07-01 至 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3272129
• 关键词: Bombycidae; DNA binding protein; Drosophilidae; Insecta; chorioallantoic membrane; developmental genetics; eukaryote; evolution; gene expression; genetic manipulation; genetic mapping; genetic promoter element; genetic regulatory element; molecular cloning; mutant; nucleic acid sequence; protein structure; tissue /cell culture; transcription factor; transfection

In recent years it has become obvious that multigene families are important features of eukaryotic genomes. Gene families are especially important for development, since their members frequently diversify after reduplication, and come to be expressed in different cells or disparate times during development. Diversified members are favorable for analysis of the molecular mechanisms that regulate gene expression in development, and of the evolution of these mechanisms. Such studies are clearly health related, insofar as correctly regulated gene expression is fundamental to proper development and health. The families of chorion (eggshell) genes of fruitflies and silkmoths exhibit a high degree of tissue, temporal and spatial regulation during development and high conservation of regulatory mechanisms during evolution, in the face of considerable structural diversification. Taking advantage of recent progress in characterizing chorion cis-regulatory elements and putative trans-acting (CF) factors, we intend to study regulatory protein-DNA interactions in an evolutionary context, using the moth A/B.L12 and the fly s15 promoters. The specific objectives are: 1. To map on the A/B.L12 promoter binding sites for fly CF factors. 2. To verify which CF components are indeed regulatory in vivo, using a combination of cell culture assays and detection of CF factors on the promoter in vivo through immunoprecipitation of chromatin. 3. To clone and characterize moth cognates of these functionally important factors. 4. To study in detail the DNA recognition properties and mechanisms of selected functionally important CF factors, from both Drosophila and the moth, with a view towards understanding the paradox of evolutionarily conserved chorion gene regulation, in the face of promoter sequence diversification.

近年来，多基因家族的重要性已变得显而易见 真核生物基因组的特征。 基因家族对于 发展，因为其成员在重复后经常多样化， 并在不同的细胞或不同的时间表达 发展。 多元化的成员有利于分析 调节发育过程中基因表达的分子机制，以及 这些机制的演变。 此类研究显然是健康的 相关，只要正确调节基因表达是基础 适当的发展和健康。 果蝇和蚕蛾的绒毛膜（蛋壳）基因家族 表现出高度的组织、时间和空间调节 监管机制的发展和高度保护 面对相当大的结构多样化，进化。 服用 绒毛膜顺式调节特征的最新进展的优势 元素和假定的反式作用（CF）因素，我们打算研究 进化背景下的调节蛋白-DNA 相互作用，使用 蛾 A/B.L12 和果蝇 s15 启动子。 具体目标是： 1. 绘制果蝇 CF 因子的 A/B.L12 启动子结合位点图。 2. 为了验证哪些 CF 成分确实在体内具有调节作用，使用 结合细胞培养测定和 CF 因子检测 通过染色质免疫沉淀在体内启动子。 3. 克隆和表征这些具有重要功能的蛾类同源物 因素。 4. 详细研究DNA识别特性和机制 从果蝇和 蛾，以期理解进化论的悖论 面对启动子序列，保守的绒毛膜基因调控 多样化。