Dear All. A question on protein expression. We have been doing small scale test expressions in 15ml of terrific broth+kanamycin using E.coli KRX cells in falcon tubes. On reaching OD600 of ~0.6 we induced with rhamnose+IPTG and expressed for 4 hours at 37 C. There is a big band corresponding to our protein in the soluble fraction and not much in the insoluble fraction. However, on scaling up to 1 L TB cultures (with same concentrations of kanamycin and inducing with same concs of rhamnose+IPTG) we don't get strong over expression. Has anyone else experienced problems when scaling up expression? (and more importantly, solved them?)
best wishes James -- Dr. James W. Murray David Phillips Research Fellow Division of Molecular Biosciences Imperial College, LONDON Tel: +44 (0)20 759 48895
