Overview of Protein Expression in Pichia pastoris

David R. Higgins1

1 Invitrogen Corporation, San Diego
Publication Name:  Current Protocols in Protein Science
Unit Number:  Unit 5.7
DOI:  10.1002/0471140864.ps0507s02
Online Posting Date:  May, 2001
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Pichia pastoris is a methylotrophic yeast and can be used as a heterologous expression system. This microorganism is as easy to manipulate as Escherichia coli, but has many of the advantages of eukaryotic expression (e.g., protein processing, folding, and postā€translational modifications), and it is faster, easier, and cheaper to use than other eukaryotic expression systems, such as baculovirus or mammalian tissue culture. It also generally yields higher expression levels. This overview discusses important considerations for the use of Pichia pastoris, including strains for expression, expression plasmids, transformation by integration, and postā€translational modifications. Examples of expression are given and finally, legal issues regarding patent rights for heterologous protein expression in Pichia pastoris are described.

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Table of Contents

  • General Characteristics of Pichia pastoris
  • Heterologous Protein Expression
  • Commentary
  • Literature Cited
  • Figures
  • Tables
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Literature Cited

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Key References
   Cregg et al., 1993. See above.
  Good overview of Pichia pastoris as an expression system with a discussion of glycosylation of expressed proteins.
   Scorer et al., 1993. See above.
  Describes in detail the advantages of intracellular versus secreted expression, the effect of multiple‐copy expression cassettes on expression levels, and an overall approach to optimizing expression levels.
   Scorer et al., 1994. See above.
  Details the generation of multiple‐copy expression cassette recombinant strains using the bacterial Kanr gene and screening for hyperresistance to G418.
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