to get protein in cytosol from inclusion bodies
Posted 11 September 2010 - 08:21 PM
anybody could tell me how to get it inside the cytosol. any suggesstions are welcomed
Posted 12 September 2010 - 04:06 AM
Posted 12 September 2010 - 04:07 AM
Posted 14 September 2010 - 10:11 PM
Posted 15 September 2010 - 04:40 AM
so you mean that the MAX Expression is at 0.05mM IPTG at 37C but is in inclusion body?
If so, I suggest you to further reduce your IPTG to 0.025mM IPTG, and keep your cultures at room temp (i assume yours would be 25C) or at 30C.
IMHO, 18-25C would be too cold for the bacteria to grow and produce efficient protein.
Also, change to rosetta strains might help. Just have to try all the possibilities.
Just my 2 cents.
p/s: using CAPS in forum makes your sentence hard to read.
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
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