How to express toxic proteins in bacteria - correction!

Dr. Peter Gegenheimer PGegen at UKans.nolospamare.edu
Fri Sep 15 19:39:47 EST 2000


On Sat, 16 Sep 2000 00:21:39, "Emir Khatipov"
<ekhatipo at NOSPAMmidway.uchicago.edu> wrote:

ð The main inconvenience for
ð me is that when the ultracentrifugation is omitted, the lysate appears
ð cloudy, and seemingly contains chelators that cause sheering of cobalt from
ð Talon resin I used to purify His6-proteins. Is there any trick to eliminate
ð ultracentrifugation and the greazy stuff?

I don't recall that we've seen such viscosity with regular (non-His-tagged)
clones. My guess is that the viscosity comes from DNA; if so, the DNA can
normally be sheared to small fragments by squirting the lysate through a 20-
to 22-gauge syringe needle, or brief sonication, or treatment with DNase I.

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