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:: 10.3109/09687688.2011.602219

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: Biomedical Research Centre
Identification Number: 10.3109/09687688.2011.602219
Depositing User: Rebecca Jones
Date Deposited: 27 Feb 2012 16:53
Last Modified: 27 Feb 2012 16:53
URI: http://shura.shu.ac.uk/id/eprint/4876

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