Purification of active Escherichia coli ribosome recycling factor (RRF) from an osmo-regulated expression system

摘要

Ribosome release factor (RRF) from Escherichia coli was overproduced from an osmo-expression vector. More than 40/100 of cell protein was RRF after 6 h of induction. A purification scheme is described that produced 50 mg of RRF from an initial culture of 2 L. The recycling time for ribosomes synthesising the tripeptide fMet-Phe-Leu in vitro in the absence of RF3 was reduced from 40 to 15 s by the addition of purified 1.5 μM RRF.