Expression and characterization of recombinant Prunus persica eEF1A. (A) Coomassie blue staining of an SDS-PAGE gel containing His6-tagged eEF1A purification fractions. Lanes 1 and 2, eluates from the Ni-nitrilotriacetic acid column, using 200 mM and 500 mM imidazole, respectively; lanes 3 and 4, flowthrough and wash fractions of the proteins from the heparin column purifications of the samples in lanes 1 and 2, respectively; lanes 5 to 8, fractions collected from the elutions performed in the presence of 150 mM, 400 mM, 600 mM, and 1,500 mM ammonium sulfate, respectively. The lane labeled L is a molecular size marker. The arrow indicates the position of recombinant eEF1A. (B) Western blot hybridization performed using an anti-His6 antibody on an SDS-PAGE gel similar to that presented in panel A. (C and D) Autoradiograms of Northwestern hybridizations performed using the fractions corresponding to the elutions performed in the presence of 400 mM and 600 mM ammonium sulfate (lanes 1 and 2, respectively), using the (−) and (+) PLMVd strands as probes, respectively.