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Eukaryotic expression: developments for structural proteomics.

A R Aricescu ; R Assenberg ; R M Bill ; D Busso ; V T Chang ; S J Davis ; A Dubrovsky ; Lena Gustafsson (Institutionen för kemi- och bioteknik) ; Kristina Hedfalk (Institutionen för kemi- och bioteknik, Molekylär bioteknik) ; U Heinemann ; I M Jones ; D Ksiazek ; C Lang ; K Maskos ; A Messerschmidt ; S Macieira ; Y Peleg ; A Perrakis ; A Poterszman ; G Schneider ; T K Sixma ; J L Sussman ; G Sutton ; N Tarboureich ; T Zeev-Ben-Mordehai ; E Yvonne Jones
Acta crystallographica. Section D, Biological crystallography (0907-4449). Vol. 62 (2006), Pt 10, p. 1114-24.
[Artikel, refereegranskad vetenskaplig]

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.

Nyckelord: Animals, Baculoviridae, genetics, Cells, Cultured, Cloning, Molecular, Eukaryotic Cells, metabolism, Gene Expression, Glycosylation, Proteomics, methods, Selenomethionine, diagnostic use, Yeasts, metabolism

Denna post skapades 2014-12-01.
CPL Pubid: 206975


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Institutioner (Chalmers)

Institutionen för kemi- och bioteknik (2005-2014)
Institutionen för kemi- och bioteknik, Molekylär bioteknik (2005-2007)


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