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. 2004 Mar 12;101(12):4059–4064. doi: 10.1073/pnas.0400554101

Fig. 4.

Fig. 4.

Time course of polypeptide synthesis in wheat germ extracts and reticulocyte lysates. (A) The Met9-AAP-LUC-AAP-LUC construct used (DNA sequence in Fig. 5B). Designations are as in Fig. 1 A except that arrowhead I is the C terminus of the Met9-AAP-LUC-AAP intermediate. (B) Wheat germ extracts (150 μl) or reticulocyte lysates (100 μl) were programmed with the indicated mRNA (W-W, Met9-AAPw-LUC-AAPw-LUC, M-W, Met9-AAPm-LUC-AAPw-LUC; and M-M, Met9-AAPm-LUC-AAPm-LUC) and incubated at 25°C with either 10 μM or 2,000 μM Arg and 10 μM each of the other amino acids. Edeine was added at 2 min, and polypeptide products were analyzed as described in Fig. 1, except that reticulocyte samples were incubated in loading buffer at 25°C for 30 min before SDS/PAGE, and 3 μl of lysate was loaded per lane. F, full-length polypeptide; I, intermediate corresponding to Met9-AAP-LUC-AAP; N, intermediate corresponding to Met9-AAP.