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Expression of soluble and membrane proteins in E. coli

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Our lab has developed a number of tools for enhancing the expression of soluble secreted protein as well as membrane proteins in bacteria. Specifically: (1) We have explored mutagenesis/screening and also host cell engineering strategies for the expression of mammalian and prokaryotic membrane proteins. This work has led to the development of simple approaches that can be used to obtain significantly increased yields of membrane proteins in E.coli. (2) Developed a high through strategy relying on flow cytometry for the isolation of mutant proteins exhibiting enhanced expression from combinatorial libraries. This latter technology has been used for the expression maturation of scFv and FAB antibody fragments.

Representative results illustrating the power of these methods will be discussed.

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Correspondence to A James Link.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution 2.0 International License (https://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Link, A.J., Jeong, K. & Georgiou, G. Expression of soluble and membrane proteins in E. coli. Microb Cell Fact 5, S15 (2006) doi:10.1186/1475-2859-5-S1-S15

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Keywords

  • Flow Cytometry
  • Membrane Protein
  • Host Cell
  • Simple Approach
  • Mutant Protein