protein expression in M15 PREP4

Ian McFarlane i.mcfarlane at icrf.icnet.uk
Tue Nov 24 12:57:02 EST 1998


Hi,

We have expressed an N-terminal His tagged protein in M15 PREP4 E. Coli.
from Qiagen. We get a nice induction with IPTG and the protein elutes from
Ni2+ columns at around 200mM imidazole. The only fly in the ointment is a
second band. The columns were loaded in 8M urea and washed clean followed
by a renaturation step. The contaminant runs about 2kD lower than our
target and the band is diffuse (appears to extend from the target protein
down).

I think it may be proteolysis products. Has anyone else out there
experienced this sort of thing with the above strain?

Thanks,

Ian Mc



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