OPTIMIZING CELL SURFACE DISPLAY IN E COLI

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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.