Eukaryotic expression: developments for structural proteomics

Acta Crystallogr D Biol Crystallogr. 2006 Oct;62(Pt 10):1114-24. doi: 10.1107/S0907444906029805. Epub 2006 Sep 19.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Cells, Cultured
  • Cloning, Molecular
  • Eukaryotic Cells / metabolism*
  • Gene Expression
  • Glycosylation
  • Proteomics / methods*
  • Selenomethionine
  • Yeasts / metabolism


  • Selenomethionine