| Literature DB >> 26431670 |
Sarah J Routledge1, Lina Mikaliunaite2, Anjana Patel2, Michelle Clare2, Stephanie P Cartwright2, Zharain Bawa2, Martin D B Wilks3, Floren Low2, David Hardy2, Alice J Rothnie2, Roslyn M Bill4.
Abstract
Historically, recombinant membrane protein production has been a major challenge meaning that many fewer membrane protein structures have been published than those of soluble proteins. However, there has been a recent, almost exponential increase in the number of membrane protein structures being deposited in the Protein Data Bank. This suggests that empirical methods are now available that can ensure the required protein supply for these difficult targets. This review focuses on methods that are available for protein production in yeast, which is an important source of recombinant eukaryotic membrane proteins. We provide an overview of approaches to optimize the expression plasmid, host cell and culture conditions, as well as the extraction and purification of functional protein for crystallization trials in preparation for structural studies.Entities:
Keywords: Cholesterol; Expression plasmid vector; Membrane proteins; Promoter; Recombinant; SMALP; Yeast
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Year: 2015 PMID: 26431670 DOI: 10.1016/j.ymeth.2015.09.027
Source DB: PubMed Journal: Methods ISSN: 1046-2023 Impact factor: 3.608