Expression and purification of recombinant eEF1Bc protein pEsc-His/Cup-FLAG-TEF4 plasmid was transformed into tef4D strain. Yeast was pre-grown overnight at 29uC in 2 ml synthetic complete dropout medium lacking histidine (SC-Hmedium) containing 2% glucose. The volume of the media was increased up to 100 ml 16 h later and copper sulfate was added to a final concentration of 50 mM for induction of protein expression. Yeast was grown to 0.8 OD 600 (,4-6 h). Then, yeast cells were harvested and broken by glass beads in a FastPrep cell disruptor followed by Flag-affinity purification of FLAG-Tef4p protein [34] . The bacterial heterologous expression and purification of His 6tagged Tef3 protein from plasmid pTKB523 was performed as described in ref: [62] using only the Ni affinity column step.