A Functional Genomics Approach for Evaluating Protein Production Pathways

该SGER提案的长期目标是研究细胞中蛋白质合成的限制，并开发增强的异源蛋白质生产系统。这将通过研究活果蝇中不同转染细胞类型的蛋白质输出来实现。绿色荧光蛋白（GFP）将是构建体标记物，并且通过昆虫器官荧光直接观察产生。IL-2，产物蛋白质，将使用组氨酸标签融合到GFP以促进蛋白质分离，并且酶切割位点连同适当的启动子沿着并入系统中。各种启动子将用于驱动不同组织中的表达。这项工作将导致高蛋白生产细胞系和/或选择果蝇生产菌株和控制蛋白生产的因素的信息，例如，启动子、分子伴侣和折叠酶等。

The long term goal of this SGER Proposal is to investigate restrictions to protein synthesis in cells and to develop enhanced heterologous protein production systems. This will be accomplished by studying the protein output of different transfected cell types in living fruit flies. Green fluorescent protein (GFP) will be the construct marker and the production will be directly observed by insect organ fluorescence. IL-2, the product protein, will be fused to the GFP using a histidine tag to facilitate protein isolation, and an enzymatic cleavage site incorporated in the system along with an appropriate promoter. Various promoters will be used to drive expression in different tissues. The work should lead to high protein-producing cell lines and/or select producing strains of Drosophila and information on the factors controlling protein production, e.g., promoters, chaperones and foldases, etc.