Protein synthesis in cell-free reticulocyte lysates on multi-hour incubation

Cell-free protein synthesis in rabbit reticulocyte lysate translation mixtures has been studied during multi-hour incubations. In an impaired lysate obtained from cells stored at 0 degrees C before lysis, and showing a low initial rate of synthesis, translation could be stimulated by a factor of 4 by including RNase inhibitor and additional ATP and GTP. In translation mixtures prepared from normal lysates, protein synthesis could be improved by approximately 50% by the addition of excess GTP. The observed increases in protein synthesis were obtained by improved maintenance of the initial rate of synthesis.