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
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