E.coli lysis no enzyme damage
behrends at plexus.uke.uni-hamburg.de
Thu Mar 1 08:30:43 EST 2001
we are getting nice soluble expression of a
80 kDa His-Tagged enzyme in E. coli. (about
10 % soluble and 90 % inclusion bodies).
Though we can measure activity when the protein
is expressed in Sf9 cells, we were not success-
ful in E. coli so far. It is not clear whether
that is due to the differences between eukaryotic
versus prokaryotic expression or whether
our Coli-lysis by sonification is too
harsh. With Sf9 cells we use a high pressure
nitrogen bomb, but we do not want to use bacteria
in that bomb.
Is digestion with lysozyme 0.1 mg / ml for 30 min on
ice sufficient for lysis of coli? And is is regarded
as a method where expressed proteins / enzymes are
left more or less intact. Is there a better method?
Thanks for your time and help
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