SEQUENCE ARRANGEMENT IN EUCARYOTIC DNA AND CHROMOSOMES

这项调查是一个案例研究如何更高的有机体 控制一个简单的开发表达模式。 这些基因 被研究的是一对紧密间隔和发散的 转录的黑腹果蝇的卵黄蛋白基因。 这些 基因具有高度分化的表达模式， 关于生物体的发展，虽然相当简单， 有几个特异性：组织，性别，在生命周期内的时间 细胞和荷尔蒙反应 项目的第一个目标 是通过组合来识别机制的组成部分 1）使用体外诱变和种系的实验 转化以鉴定必需的序列和/或 对于正常表达模式的各方面而言足够;以及2） 使用DNA结合试验和无细胞- 转录以识别和表征蛋白质和DNA 影响YP表达模式的序列。 第二 目的是研究鉴定的DNA之间的相互作用， 件. 这将通过操作组件来完成 数量，安排，和种类，然后测试结果 通过生殖系转化、无细胞转录和物理转化， 蛋白质-DNA相互作用的研究。 第三个目标是 确定顺式作用DNA的发育特异性 控制基因的反式元件， yp 1 -2转录模式的一个方面。

This investigation is a case study of how a higher organism controls a simple development expression pattern. The genes to be studied are the pair of closely spaced and divergently transcribed yolk protein genes of Drosophila melanogaster. These genes have an expression pattern that is highly differentiated with respect to development of the organism and, though fairly simple, has several specificities: tissue, sex, timing within the lifetime of a cell, and hormonal response. The first objective of the project is to identify components of the mechanism by a combination of: 1) experiments which use in vitro mutagenesis and germ line transformation to identify sequences that are necessary and/or sufficient for aspects of the normal expression pattern; and 2) experiments which use DNA binding assays and cell free- transcription to identify and characterize proteins and DNA sequences which influence the yp expression pattern. The second objective is to examine interactions between identified DNA components. This will be done by manipulating component number, arrangement, and kind and then testing the consequences by germ line transformation, cell free transcription, and physical studies of protein-DNA interactions. The third objective is to determine the developmental specificities of cis-acting DNA elements which control a gene that acts in trans to determine one aspect of the yp1-2 transcription pattern.