Proteins in E. coli inclusion bodies

verwaerde at mbcl.rutgers.edu verwaerde at mbcl.rutgers.edu
Mon Jun 8 15:51:06 EST 1992


In article <1992May19.165336.10690 at athena.cs.uga.edu>, russell at dogwood.botany.uga.edu writes:
> When E. coli dumps a eucaryotic protein into an inclusion body,
> What is happening?
> Why has the protein become insoluble?
> 
> 
> We are trying to purify our favorite protein from an E. coli
> vector, where the protein is expressed and placed into an inclusion
> body.
> We have used guanidine HCl to resuspend the protein, and, after
> various treatments, can get enzyme activity.
> 
> Methods that we used to purify the protein originally, however,
> do not always work with this protein from the inclusion body,
> even though the prep does have enzyme activity.
> 
> Thus, we wondered how the protein might be modified in E. coli,
> or if carbohydrates or some other entangling molecule might
> exist in the inclusion body, so that we can't use the same
> purification methods.
> 
> Thanks for any help.

You're lucky in a way because your protein is insoluble you may take advantage
of it: in biotechniques (1991) vol 11 No^ p748-752 yo'll find a nice and 
gentle technique to purify any insoluble protein in inclusion bodies.


Phil Verwaerde, CABM, Rutgers University

> Russell Malmberg, russell at dogwood.botany.uga.edu



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