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REGULATION AND EVOLUTION OF MULTIGENE FAMILIES

多基因家族的调控和进化

基本信息

批准号：
2174222
负责人：
FOTIS C KAFATOS
金额：
$ 21.2万
依托单位：
HARVARD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1977
资助国家：
美国
起止时间：
1977-07-01 至 1994-11-30
项目状态：
已结题

项目摘要

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越来越明显真核生物基因组的特征。基因家族对于发展，因为它们的成员经常在重叠后多样化，并在不同的细胞或不同的时间表达，发展多样化的成员有利于分析在发育过程中调节基因表达的分子机制，这些机制的演变。这样的研究显然是健康的相关的，只要正确调控基因表达是至关重要的，适当的发展和健康。果蝇和蚕蛾的卵壳基因家族表现出高度的组织、时间和空间调节，发展和高度保守的监管机制，在结构多样化的背景下，以最近在表征绒毛膜顺式调节方面取得的进展的优势元素和假定的反式作用（CF）因子，我们打算研究调节蛋白质-DNA相互作用在进化的背景下，使用蛾A/B.L12和蝇s15启动子。具体目标是： 1.目的定位果蝇CF因子A/B.L12启动子结合位点。 2.为了验证哪些CF组分确实是体内调节性的，使用细胞培养试验和CF因子检测的组合，通过染色质的免疫沉淀在体内表达启动子。 3.克隆和鉴定这些功能重要的蛾同源物因素 4.详细研究了DNA识别的性质和机理，选择功能重要的CF因子，从果蝇和蛾，以了解进化的悖论，保守的绒毛膜基因调控，面对启动子序列多样化经营。