Molecular Genetics of the PAS Region

胚胎决定和基因表达差异模式的建立和维持对正常动物发育至关重要。在果蝇中，至少在一定程度上，胚胎的决定是由双胸和触须复合体的选择基因控制的。我一直在研究一个被称为PAS-R的遗传区域，它包含了许多相关的基因。这些基因的正常功能是正常发育所必需的，它似乎是选择基因的适当调节所必需的。本提案包括PAS-R的分子和发育遗传分析。我建议使用经典遗传学、体外诱变和种系转化来严格识别具有功能基因的单个蛋白质和rna。我将对来自这些基因的cDNA克隆进行测序，以确定它们编码的蛋白质的氨基酸序列。这可能会告诉我们一些关于它们的功能，并且是为蛋白质制作抗体探针的有价值的信息。我建议使用发育和分子方法来鉴定需要PAS-R基因产物存在的调控基因。将开发实验系统，使我们能够研究这种调节，并确定PAS-R蛋白的功能域和目标识别序列。导致成人发育的基因表达的编排在很大程度上仍然不为人所知。深入了解基因的功能，如在PAS-R区域发现的那些被认为调节已知的发育必需基因的表达的基因，应该从这项研究中得出。

The establishment and maintenance of embryonic determination and differential patterns of gene expression is essential for normal animal development. In Drosophila embryonic determination is, at least in part, under the control of the selector genes of the bithorax and Antennapedia complexes. I have been studying a genetic region, referred to as the PAS-R, that contains a number of related genes. The proper function of these genes is essential for normal development and it appears to be required for the proper regulation of selector genes. This proposal consists of a molecular and developmental genetic analysis of the PAS-R. I propose to use classical genetics, in vitro mutagenesis, and germ line transformation to rigorously identify individual proteins and RNAs with functional genes. I will sequence cDNA clones derived from these genes to determine the amino acid sequences of the proteins they encode. This may tell us something about their function, and is valuable information for making antibody probes for the proteins. I propose to use both developmental and molecular approaches to identify genes whose regulation requires the presence of PAS-R gene products. Experimental systems will be developed that will enable us to study this regulation, and to identify functional domains of PAS-R proteins and target recognition sequences. The orchestration of gene expression which results in the development of an adult is still largely not understood. Insight into the function of genes such as those found in the PAS-R region which are thought to regulate the expression of genes known to be developmentally essential should result from this study.