APTAMERS TO ANALYZE REGULATORY NETWORKS IN DROSOPHILA

这个项目的目标是开发肽适体的用途 在体内干扰蛋白质功能。肽适体短 肽（与平台分子融合，E.大肠杆菌硫氧还蛋白）， 特别是靶蛋白。破坏特定蛋白质的适体 可以使用酵母双杂交系统选择蛋白质相互作用，然后 在果蝇中以空间和时间控制的模式表达。 这个小组以前的结果表明，肽适体可以用于 特异性靶向和抑制体内细胞周期调节因子，并使用两种 混合方法，他们已经确定了一个蛋白质相互作用网络， 包括已知的和潜在的细胞周期调节剂。该提案有四个 具体目标。1)为了改进分离和亚克隆的技术， 适体2)为了测试aptomer方法，通过表达破坏 果蝇眼睛中Cdk2、Cyclin E和Dacapo之间的相互作用。3)到 从相互作用的蛋白质网络中部分表征新蛋白质， 确定一个可能调节细胞周期的核心组， 发展中的眼睛4)利用肽适体分析这个核心的功能 一组蛋白质。

The goal of this project is to develop the use of peptide aptomers that interfere with protein function in vivo. Peptide aptomers are short peptides (fused to a platform molecule, E. coli thioredoxin) that interact specifically with a target protein. Aptomers that disrupt specific protein/ protein interactions can be selected using the yeast two-hybrid system and then expressed in Drosophila in spatially and temporally controlled patterns. Previous results from this group have shown that peptide aptomers can be used to specifically target and inhibit cell cycle regulators in vivo, and using two hybrid approaches they have identified a network of protein interactors, that includes known and potential cell cycle regulators. The proposal has four Specific Aims. 1) To improve the technology for the isolation and subcloning of aptomers. 2) To test the aptomer approach by expressing aptomers that disrupt interactions between Cdk2, Cyclin E and Dacapo in the Drosophila eye. 3) To partially characterize novel proteins from a network of interacting proteins to identify a core group that are likely to regulate the cell cycle in the developing eye. 4) To use peptide aptomers to analyze the function of this core group of proteins.