Yeast expression

Yeast is an organism which is well-suited for the expression of recombinant proteins or the production of endogenous complexes

 
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Yeast expression Details

Yeast cells constitute a convenient alternative host for production of either recombinant eukaryotic proteins or endogenous complexes. Advantages include the low cost of growing material, robustness of the system, easy genetic manipulation, and the presence of endogenous eukaryotic-type protein modification and chaperone systems

User Guide

Overexpression of recombinant protein in yeast benefits from the absence of size limitation and the possibility to overproduce several factors simultaneously.

Overexpression of genes of interest in Saccharomyces cerevisiae is easy to set up provided that the coding sequence(s) is (are) fused to appropriate control regions (promoter, regulators…). While users have the choice of many parameters (e.g., regulated versus constitutive promoters, plasmidic versus chromosomal expression…), the preferred system involves overexpression driven by a galactose regulated strong promoter from a multicopy plasmid. The procedure consists then in the construction of the expression vector, chemical yeast transformation and culture under a selective medium.

Because protein yields can be lower in yeast than in a bacterial system, efficient purification strategies, such as TAP method, may be required. The TAP tag may be inserted in a plasmid driving protein overexpression or can be targeted to purify endogenous protein complexes.

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