Overexpression and purification of elongation factor 3 from Saccharomyces cerevisiae

The translational elongation factor 3 (EF-3) from Saccharomyces cerevisiae was overexpressed and purified to near homogeneity from the post-ribosomal supernatant fraction (S-100). A detailed protocol for the isolation of overexpressed EF-3 is presented. The procedure involves ion-exchange chromatography on DEAE-Sepharose and CM-Sepharose and affinity chromatography on ATP-agarose. A protein purity of > or = 96% was established by quantitating the silver-stained SDS/polyacrylamide gel. The present method facilitates isolation of EF-3 in large amounts in high yield.