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Eukaryotic expression: developments for structural proteomics

Item Type:Article
Title:Eukaryotic expression: developments for structural proteomics
Creators Name:Aricescu, A.R. and Assenberg, R. and Bill, R.M. and Busso, D. and Chang, V.T. and Davis, S.J. and Dubrovsky, A. and Gustafsson, L. and Hedfalk, K. and Heinemann, U. and Jones, I.M. and Ksiazek, D. and Lang, C. and Maskos, K. and Messerschmidt, A. and Macieira, S. and Peleg, Y. and Perrakis, A. and Poterszman, A. and Schneider, G. and Sixma, T.K. and Sussman, J.L. and Sutton, G. and Tarboureich, N. and Zeev-Ben-Mordehai, T. and Jones, E.Y.
Abstract:The production of sufficient quantities of protein is an essential prelude to a structure determination, but for many viral and human proteins this cannot be achieved using prokaryotic expression systems. Groups in the Structural Proteomics In Europe (SPINE) consortium have developed and implemented high-throughput (HTP) methodologies for cloning, expression screening and protein production in eukaryotic systems. Studies focused on three systems: yeast (Pichia pastoris and Saccharomyces cerevisiae), baculovirus-infected insect cells and transient expression in mammalian cells. Suitable vectors for HTP cloning are described and results from their use in expression screening and protein-production pipelines are reported. Strategies for co-expression, selenomethionine labelling (in all three eukaryotic systems) and control of glycosylation (for secreted proteins in mammalian cells) are assessed.
Keywords:Baculoviridae, Cultured Cells, Eukaryotic Cells, Gene Expression, Glycosylation, Molecular Cloning, Proteomics, Selenomethionine, Yeasts, Animals
Source:Acta Crystallographica Section D
ISSN:0907-4449
Publisher:International Union of Crystallography (U.K.)
Volume:62
Number:Pt. 10
Page Range:1114-1124
Date:October 2006
Official Publication:https://doi.org/10.1107/S0907444906029805
PubMed:View item in PubMed

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