Cloning and Multi-Subunit Expression of Mitochondrial Membrane Protein Complexes in Saccharomyces cerevisiae
- 448 Downloads
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 wordspBEVY Saccharomyces cerevisiae Multi-subunit expression Mitochondrial membrane protein LiAc/SS transformation Membrane protein complexes
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.
- 5.Miller CA, Martinat MA, Hyman LE (1998) Assessment of aryl hydrocarbon receptor complex interactions using pBEVY plasmids expression vectors with bi-directional promoters for use in Saccharomyces cerevisiae. Nucleic Acids Res 26(15):3577–3583. https://doi.org/10.1093/nar/26.15.3577CrossRefPubMedGoogle Scholar