Preparation of cell-free extracts for protein purification. A
1 litre baffled Erlenmeyer flask containing 400 ml of LB medium
was inoculated with 10 ml of an overnight culture grown on LB
kanamycin medium. Cells were grown at 37 uC with shaking at
250 r.p.m. using an Innova I2400 incubator shaker (New Brunswick
Scientific). When cells reached an OD600 of 0.6–0.8 the temperature
was reduced to 30 uC, and protein production was induced by
addition of IPTG to 0.5 mM. Six hours after induction, cells were