ACCUMULATION OF TRANSLATIONAL INHIBITOR DURING MULTI-HOUR CELL-FREE PROTEIN-SYNTHESIS REACTION USING RABBIT RETICULOCYTE LYSATE

A cell-free protein synthesis in rabbit reticulocyte lysates stops wit hin 2 h, which limits the productivity of the cell-free system. Here w e show that the accumulation of a translational inhibitor liberated fr om the post-ribosomal supernatant during the reaction time is responsi ble for the halt in the translational reaction. Since EGTA was found t o suppress the effect of the inhibitor and the absorbance of a Ca2+ in dicator was observed to increase, Ca2+ is suggested to be the inhibito r that accumulates in reticulocyte lysates during the cell-free protei n synthesis reaction.