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.

metabolism

diagnostic use

Yeasts

Eukaryotic Cells

Gene Expression

Cells

Cultured

methods

Glycosylation

Proteomics

Cloning

Animals

genetics

metabolism

Baculoviridae

Molecular

Selenomethionine

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-institutet

C Lang

Technische Universitat Berlin

K Maskos

Max Planck-institutet

A Messerschmidt

Max Planck-institutet

S Macieira

Max Planck-institutet

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 1114-24

Ämneskategorier

Biologiska vetenskaper

DOI

10.1107/S0907444906029805

PubMed

17001089