NOTE: In the version of this Review initially published, an author (B. Martin Hallberg) was left off of the author list. This information has been added to the HTML and PDF versions of the Review.
In selecting a method to produce a recombinant protein, a researcher is faced with a bewildering array of choices as to where to start. To facilitate decision-making, we describe a consensus 'what to try first' strategy based on our collective analysis of the expression and purification of over 10,000 different proteins. This review presents methods that could be applied at the outset of any project, a prioritized list of alternate strategies and a list of pitfalls that trip many new investigators.
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The Structural Genomics Consortium is a registered charity (number 1097737) that receives funds from the Canadian Institutes for Health Research, the Canadian Foundation for Innovation, Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, Karolinska Institutet, the Knut and Alice Wallenberg Foundation, the Ontario Innovation Trust, the Ontario Ministry for Research and Innovation, Merck & Co., Inc., the Novartis Research Foundation, the Swedish Agency for Innovation Systems, the Swedish Foundation for Strategic Research and the Wellcome Trust. The New York Structural GenomiX Research Center for Structural Genomics is supported by the US National Institute of General Medical Sciences (U54 GM074945). Work at the MDC was supported by the German Federal Ministry for Education and Research (BMBF) through the Leitprojektverbund Proteinstrukturfabrik and the German National Genome Network (NGFN; FKZ 01GR0471, 01GR0472), and by the Fonds der Chemischen Industrie. The Protein Sample Production Facility is funded by the Helmholtz Association of German Research Centres. The China Structural Genomics Consortium is supported by the National 863 Hi-Tech Research and Development Program of China. The Israel Structural Proteomics Center is supported by The Israel Ministry of Science, Culture and Sport, the Divadol Foundation, the Neuman Foundation, the European Commission Sixth Framework Research and Technological Development Programme 'SPINE2-Complexes' Project under contract 031220. The RIKEN Structural Genomics/Proteomics Initiative was supported by the National Project on Protein Structural and Functional Analyses, Ministry of Education, Culture, Sports, Science and Technology of Japan. The Joint Center for Structural Genomics is supported by the US National Institutes of Health (NIH) Protein Structure Initiative grant U54 GM074898 from the NIGMS. The Northeast Structural Genomics Consortium is supported by the NIH NIGMS (U54-GM074958). The Midwest Center for Structural Genomics is supported by the NIH (GM074942) and by the US Department of Energy, Office of Biological and Environmental Research (DE-AC02-06CH11357). The Oxford Protein Production Facility is funded by the UK Medical Research Council and Biotechnology and Biological Sciences Research Council. SPINE2-Complexes is funded by the European Commission (contract 031220) under the Framework 6 RTD Programme and is coordinated from the Division of Structural Biology, Wellcome Trust Centre for Human Genetics, Oxford, UK. The Berkeley Structural Genomics Center is supported by the NIH (GM62412). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIGMS or the NIH.
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Structural Genomics Consortium., Gräslund, S., Nordlund, P. et al. Protein production and purification. Nat Methods 5, 135–146 (2008). https://doi.org/10.1038/nmeth.f.202
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