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Expression and -N end rule

Expression and -N end rule - Protocols and Methods Forum

Expression and -N end rule - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 04-22-2009, 10:10 PM
David-Paul Minde
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Default Expression and -N end rule



.... maybe, you even find a nearby colleague who is (as me) using a
non-commercial SUMO system which does the exact same good job for
overexpression and solubility in E. coli
SUMO system is really fun to use (if even more simple approaches like
His-tag in a pET vector dont work). Great (3D) specificity, ultrafast
cleavage and easy mass production of the corresponding enzyme for tag
removal (Ulp1; we got 150 mg from 200 ml cells - enough for removing tag
from some 37.5 g of fusion proteins which is probably more than the whole
lab will need for the last couple years of my PhD
The literature on dramatic enhancements with codon optimized gene synthesis
is very,very thin. So you better check if there are indeed extreme codons
biases in your target before investing in that approach. On the other hand
it might save labour cost in cloning work in your group and be cost-neutral
for your small target.
cheers,
David

Science is what happens while we are making other plans (~John Lennon)
(PS
I am currently working in the netherlands. Really a public sport here to
save money
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