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Mitosis pp 63-80 | Cite as

Production of Mitotic Regulators Using an Autoselection System for Protein Expression in Budding Yeast

  • Marco Geymonat
  • Adonis Spanos
  • Steven Sedgwick
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 545)

Abstract

A novel protein expression system in budding yeast is described which has been used to express many yeast mitotic regulators as well as a wide range of other recombinant proteins from several different species. The expression system relies on autoselection with essential genes to maintain high copy numbers of expression plasmids. Autoselection permits expression cells to be grown in rich medium with no need for plasmid selection with drugs or nutritional conditions. This optimizes growth and expression of recombinant proteins. The use of the expression system is illustrated by purifying budding yeast mitotic regulators, Cdc14 and Net1, and recapitulating their activities in vitro.

Key words

Protein expression budding yeast recombinant proteins autoselection 

References

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Marco Geymonat
    • 1
  • Adonis Spanos
    • 1
  • Steven Sedgwick
    • 1
  1. 1.Stem Cell Biology and Developmental GeneticsNational Institute for Medical ResearchUK

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