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Cloning and Multi-Subunit Expression of Mitochondrial Membrane Protein Complexes in Saccharomyces cerevisiae

  • Porsha L. R. Shaw
  • Kathryn A. Diederichs
  • Ashley Pitt
  • Sarah E. Rollauer
  • Susan K. BuchananEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2127)

Abstract

Saccharomyces cerevisiae is a useful eukaryotic expression system for mitochondrial membrane proteins due to its ease of growth and ability to provide a native membrane environment. The development of the pBEVY vector system has further increased the potential of S. cerevisiae as an expression system by creating a method for expressing multiple proteins simultaneously. This vector system is amenable to the expression and purification of multi-subunit protein complexes. Here we describe the cloning, yeast transformation, and co-expression of multi-subunit outer mitochondrial membrane complexes using the pBEVY vector system.

Key words

pBEVY Saccharomyces cerevisiae Multi-subunit expression Mitochondrial membrane protein LiAc/SS transformation Membrane protein complexes 

Notes

Acknowledgments

The authors would like to thank Dr. Edmund Kunji (Cambridge, UK) for the yeast expression strain and the pBEVY vectors. This work was supported by the Intramural Research Program of the NIH, the National Institute of Diabetes & Digestive & Kidney Diseases.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Porsha L. R. Shaw
    • 1
  • Kathryn A. Diederichs
    • 1
  • Ashley Pitt
    • 1
  • Sarah E. Rollauer
    • 1
    • 2
  • Susan K. Buchanan
    • 1
    Email author
  1. 1.Laboratory of Molecular BiologyNational Institute of Diabetes & Digestive & Kidney Diseases, National Institutes of HealthBethesdaUSA
  2. 2.Vertex PharmaceuticalsBostonUSA

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