Eukaryotic expression: developments for structural proteomics.
Artikel i vetenskaplig tidskrift, 2006
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.
Baculoviridae
Cells
Cultured
methods
Cloning
diagnostic use
genetics
Animals
Eukaryotic Cells
Glycosylation
Gene Expression
Selenomethionine
Yeasts
metabolism
Proteomics
Molecular
metabolism
Författare
A R Aricescu
Wellcome Trust Centre for Human Genetics
R Assenberg
Wellcome Trust Centre for Human Genetics
Joakim Norbeck
Aston University
Chalmers University of Technology
D Busso
IGBMC Institut de Genetique et de Biologie Moleculaire et Cellulaire
V T Chang
John Radcliffe Hospital
S J Davis
John Radcliffe Hospital
A Dubrovsky
Karolinska Institutet
Lena Gustafsson
Chalmers, Kemi- och bioteknik
Kristina Hedfalk
Chalmers, Kemi- och bioteknik, Molekylär bioteknik
U Heinemann
Max Delbruck Center for Molecular Medicine
I M Jones
University of Reading
D Ksiazek
Max Planck Institute of Biochemistry
C Lang
Technische Universitat Berlin
K Maskos
Max Planck Institute of Biochemistry
A Messerschmidt
Max Planck Institute of Biochemistry
S Macieira
Max Planck Institute of Biochemistry
Y Peleg
Weizmann Institute of Science Israel
A Perrakis
The Netherlands Cancer Institute
A Poterszman
IGBMC Institut de Genetique et de Biologie Moleculaire et Cellulaire
G Schneider
John Radcliffe Hospital
T K Sixma
The Netherlands Cancer Institute
J L Sussman
Weizmann Institute of Science Israel
G Sutton
Wellcome Trust Centre for Human Genetics
N Tarboureich
Institut Laue-Langevin
T Zeev-Ben-Mordehai
Weizmann Institute of Science Israel
E Yvonne Jones
Wellcome Trust Centre for Human Genetics
Acta Crystallographica Section D: Biological Crystallography
0907-4449 (ISSN) 1399-0047 (eISSN)
Vol. 62 Pt 10 1114-24Ämneskategorier (SSIF 2011)
Biologiska vetenskaper
DOI
10.1107/S0907444906029805
PubMed
17001089