Unit

UNIT 13.2 Expression of Recombinant Genes in the Yeast Pichia pastoris

  1. James M. Cregg1,2,
  2. Ilya Tolstorukov1,2,
  3. Anasua Kusari1,
  4. Anthony Jay Sunga1,
  5. Knut Madden2,
  6. Thomas Chappell2

Published Online: 1 DEC 2010

DOI: 10.1002/9780470089941.et1302s04

Current Protocols Essential Laboratory Techniques

Current Protocols Essential Laboratory Techniques

How to Cite

Cregg, J. M., Tolstorukov, I., Kusari, A., Sunga, A. J., Madden, K. and Chappell, T. 2010. Expression of Recombinant Genes in the Yeast Pichia pastoris. Current Protocols Essential Laboratory Techniques. 4:13.2:13.2.1–13.2.14.

Author Information

  1. 1

    Keck Graduate Institute of Applied Life Sciences, Claremont, California

  2. 2

    Biogrammatics, Inc., Carlsbad, California

Publication History

  1. Published Online: 1 DEC 2010
  2. Published Print: DEC 2010

Abstract

The synthesis of specific recombinant proteins using single-celled organisms from bacteria to mammalian tissue culture cells has become a major source of biopharmaceutical products for the industry and a source of a wide variety of proteins for academic research. A range of organisms are utilized for this purpose. One of the newest and most promising of these is the yeast Pichia pastoris. This article provides detailed basic protocols for the expression of heterologous genes and the synthesis of recombinant proteins utilizing this yeast. Specifically provided are protocols for the insertion of foreign vector DNAs into the yeast by electroporation, amplification of vector sequences by the post-translational vector amplification (PTVA) method, and growth and expression of foreign genes in shake flask cultures. Curr. Protoc. Essential Lab. Tech. 4:13.2.1-13.2.14. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • recombinant proteins;
  • foreign gene expression;
  • heterologous genes