Team:Berkeley Wetlab/Project Overview

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Cell surface display requires that a protein of interest be exposed to the extracellular environment but remain anchored to the outer membrane.  This is done by the fusion of a domain of interest, referred to as the passenger, to a protein, referred to as the displayer, that naturally inserts itself into the outermembrane.  Genetic devices for cell surface display can be thought of as composed of three basic components: the passenger domain that will be displayed to the extracellular environment, the displayer domain which will anchor the passenger to the outer membrane, and the structural spacer elements that link these two regions.<br><br>
Cell surface display requires that a protein of interest be exposed to the extracellular environment but remain anchored to the outer membrane.  This is done by the fusion of a domain of interest, referred to as the passenger, to a protein, referred to as the displayer, that naturally inserts itself into the outermembrane.  Genetic devices for cell surface display can be thought of as composed of three basic components: the passenger domain that will be displayed to the extracellular environment, the displayer domain which will anchor the passenger to the outer membrane, and the structural spacer elements that link these two regions.<br><br>
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While it is almost certain that for a given passenger, a combination of displayer and structural spacers exists that leads to functional display, it is not clear what this combination is or how to chose such a combination rationally.  We therefore employed a combinatorial approach, testing many variations of display and spacers for each passenger.  This data allows future investigators to estimate the number of combinations that must be constructed in order to find functional display, and helps guide design to types of combinations which are more likely to be successful.
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While it is almost certain that for a given passenger, a combination of displayer and structural spacers exists that leads to functional display, it is not clear what this combination is or how to chose such a combination rationally.  We therefore employed a combinatorial approach, testing many variations of display and spacers for each passenger.  This data allows future investigators to estimate the number of combinations that must be constructed in order to find functional display, and helps investigators chose subsets of combinations within the design space that are most likely to yield success.

Revision as of 22:07, 18 October 2009

Schematic.jpg

Cell surface display requires that a protein of interest be exposed to the extracellular environment but remain anchored to the outer membrane. This is done by the fusion of a domain of interest, referred to as the passenger, to a protein, referred to as the displayer, that naturally inserts itself into the outermembrane. Genetic devices for cell surface display can be thought of as composed of three basic components: the passenger domain that will be displayed to the extracellular environment, the displayer domain which will anchor the passenger to the outer membrane, and the structural spacer elements that link these two regions.

While it is almost certain that for a given passenger, a combination of displayer and structural spacers exists that leads to functional display, it is not clear what this combination is or how to chose such a combination rationally. We therefore employed a combinatorial approach, testing many variations of display and spacers for each passenger. This data allows future investigators to estimate the number of combinations that must be constructed in order to find functional display, and helps investigators chose subsets of combinations within the design space that are most likely to yield success.