OPTIMIZING CELL SURFACE DISPLAY IN E COLI

描述：外源蛋白在细胞外膜上的表面展示。 细菌细胞提供了一种独特的和潜在的强大的手段， for novel新functionality功能.然而，实际上， 这种技术的实现阻碍了其广泛应用。 用于展示的最佳表征的锚蛋白之一是融合蛋白 大调E的一个片段大肠杆菌脂蛋白（Lpp）和一个片段 外膜蛋白OmpA科尔曼博士建议使用 Lpp-OmpA与绿色荧光蛋白的融合（Lpp-OmpA-GFP）， 帮助设计一种在大肠杆菌中更稳定膜锚蛋白。大肠杆菌， 具有最佳的表面表达水平，允许多轮 细胞分选，并允许细菌生长到更高的细胞 密度和温度更高。一种改进的细胞表面显示器， 系统将允许更多种类的蛋白质在 更易于管理的条件，为促进 抗体开发和酶进化等多种技术。

DESCRIPTION: Surface display of foreign proteins on the outer membrane of bacterial cells provides a unique and potentially powerful means to screen for novel functionality. However, practical constraints on the implementation of this technology prevents its widespread application. One of the best characterized anchor proteins for display is a fusion between a fragment of the major E. coli lipoprotein (Lpp) and a fragment of the outer membrane protein, OmpA. Dr. Coleman proposes to use a fusion of Lpp-OmpA with the Green Fluorescent Protein (Lpp-OmpA-GFP) to help engineer a membrane anchor protein that is more stable in E. coli, has the optimum level of surface expression, allows multiple rounds of cell sorting, and which permits the bacteria to be grown to a higher cell density and at a higher temperature. An improved cell surface display system will allow a greater variety of proteins to be expressed under more manageable conditions, providing an impetus for the advancement of such diverse technologies as antibody development and enzyme evolution.