Improving recombinant eukaryotic membrane protein yields inPichia pastoris: the importance of codon optimization and clone selection

ÖBERG, Fredrik, SJÖHAMN, Jennie, CONNER, Matthew, BILL, Roslyn M. and HEDFALK, Kristina (2011). Improving recombinant eukaryotic membrane protein yields inPichia pastoris: the importance of codon optimization and clone selection. Molecular Membrane Biology, 28 (6), 398-411.

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Link to published version:: https://doi.org/10.3109/09687688.2011.602219
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    Abstract

    In the last 15 years, 80% of all recombinant proteins reported in the literature were produced in the bacterium, Escherichia coli, or the yeast, Pichia pastoris. Nonetheless, developing effective general strategies for producing recombinant eukaryotic membrane proteins in these organisms remains a particular challenge. Using a validated screening procedure together with accurate yield quantitation, we therefore wished to establish the critical steps contributing to high yields of recombinant eukaryotic membrane protein in P. pastoris. Whilst the use of fusion partners to generate chimeric constructs and directed mutagenesis have previously been shown to be effective in bacterial hosts, we conclude that this approach is not transferable to yeast. Rather, codon optimization and the preparation and selection of high-yielding P. pastoris clones are effective strategies for maximizing yields of human aquaporins. Read More: http://informahealthcare.com/doi/abs/10.3109/09687688.2011.602219

    Item Type: Article
    Research Institute, Centre or Group - Does NOT include content added after October 2018: Biomolecular Sciences Research Centre
    Identification Number: https://doi.org/10.3109/09687688.2011.602219
    Page Range: 398-411
    Depositing User: Rebecca Jones
    Date Deposited: 27 Feb 2012 16:53
    Last Modified: 09 Oct 2018 13:03
    URI: http://shura.shu.ac.uk/id/eprint/4876

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