Purification of wheat factors involving in translation initiation for reconstitution of protein synthesis

Abstract

We are making efforts to construct a reconstituted cell-free protein synthesis system with the components from wheat. We report here on our progress in purification of some factors expected to be needed for reconstitution of efficient translation initiation. eIF1A and eIF5 were overexpressed in E. coli and purified essentially according to the literature. Some hydrophobic chromatography media were tested if they can be used for purification of the wheat ribosomes. We also fractionated a wheat embryo extract and analyzed which fractions contain an RNA binding protein that bind to a translational enhancer sequence.