soluble expression of GST-fusion protein

yangx yangx at PUBLIC.EAST.CN.NET
Sun Jan 19 17:11:01 EST 1997


Dear Sir:

I just do research on purification of GST fusion protein, and have some
 problems, I wonder if you would like to help me.

Using the pGEX-5x-1 vector, I fused a cytokine with GST, and expression
it in E.coli, the expression level is about 40% of the whole bacterial
protein, but it formed inclusion body, the recovery rate is extremely
low,
though I had changed the conditions of recovery in many ways, I would 
like to know if the fusion protein can be expressed in soluble form if
possible, how did I change the condition of expression, if impossible, 
how did I recover the fusion protein? in addition, there are 6 cystine
in
the sequence of cytokine, can I get the active cytokine from solvent
inclusion body by GST affinity chromatography?

Thank you very much for your consideration, I look forward to hearing
from you.
 
Beat wishes!

Xiaolin Wu  Ph.D.



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