GST fusion protein solubility

atanasio atanasio at GUGU.USAL.ES
Tue Jan 13 07:41:46 EST 1998


Hi,

I am experiencing troubles with GST fusion proteins (total Mr 65000). 
Most of the protein stays in the inclusion body pellet. It can be 
solubilized by 8M urea, but thereafter the fusion protein will not 
efficiently bind to GSH-agarose. We have tried to grow bacteria at RT, 
but with limited success. We have also tried to make a MBP fusion but we 
are experiencing the same troubles.

Does anybody now to how to make the fusion proteins soluble without 
interfering with their ability to interact with the affinity column?. 
Do you think it is worth to shift to poly-His tagging?.

Thanks for all, and best of luck to the readers.


Atanasio Pandiella
Instituto de Microbiología Bioquímica
Edificio Departamental
Avda. del Campo Charro s/n
37007-Salamanca-Spain

Phone: +34-23-120561
fax: +34-23-224876
e-mail: atanasio at gugu.usal.es



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