Trans-Regulators of Chorion Gene Expression During Development

Kafatos博士建议继续分析蛾子绒毛膜启动子的顺式调控元件，并通过生化分级、分子克隆和体外转录试验的功能分析来表征反式因子。具体目标如下：1)利用LacZ报告基因，详细确定转入果蝇基因组的蛾绒毛膜基因在卵泡特异性调控中所需的元件。2)从果蝇和蚕蛾中分离、纯化和鉴定可能的反式因子。3)研究时间规划中不同绒毛膜基因的这些因素的相对亲和力。4)反式因子序列的分子克隆。5)建立体外转录系统，以检测和确认纯化或克隆的因子的性质。6)利用因子克隆研究绒毛的调节，如绒毛形成过程中因子的亲和力和浓度的变化，以及它们在其他组织和发育阶段的分布。绒毛膜蛋白，用于形成飞蛾和苍蝇的蛋壳。它们的合成为研究复杂的多基因家族的发育调控表达提供了一个很好的系统。

Dr. Kafatos proposes to continue an analysis of cis-regulatory elements of the moth chorion promoters, and to characterize the trans factors by biochemical fractionation, molecular cloning, and functional analysis with in vitro transcription assays. Specific objectives are as follows: 1) Use a reporter lacZ gene to determine in detail elements that are necessary for follicle specific regulation of moth chorion genes transferred into the Drosophila genome. 2) Biochemically fractionate, purify and characterize putative trans-factors from Drosophila and silkmoths. 3) Study the relative affinities of these factors for chorion genes that differ in temporal programming. 4)Molecular cloning of trans-factor sequences. 5) Development of an in vitro transcription system, to assay and confirm the nature of purified or cloned factors. 6) Use of the factor clones to study chorion regulation, e.g. affinities and changes in concentration of the factors during choriogenesis, and their distribution in other tissues and developmental stages. %%% The chorion proteins and used to form the eggshell of the moth and fly. Their synthesis represents an excellent system in which to study the developmentally regulated expression of a complex, multigene family.