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Abstract:
Polypeptide elongation factor EF-Tu can be isolated from bacterial cell extracts in two fractionation steps. The first is ion-exchange chromatography on DEAE-Sepharose, CL-6B, and the second is gel filtration on AcA 44. The method is illustrated with extracts from Escherichia coli, Bacillus stearothermophilus, and the thermophilic bacterium PS3. The extracts were obtained from lysozyme-treated cells and were processed without high-speed centrifugation or ammonium sulfate fractionation. The procedure is simple and rapid, gives higher yields than previous methods, and is easily scaled to any size preparation. The procedure also produces fractions enriched in the other polypeptide elongation factors EF-Ts and EF-G.