Initiation any GST-fusion protein and His-tag protein purification?

[Cheng Luo]

Actually I didn't intend to send the content. 

I have a GST-X fusion protein to be purified and crysatlized. The protein is 
very unstable after cleavage with thrombin,  can be only stable in PMSF in 
pH8.0 for about 1 month, after concnetrated to 10mG/ml it is easy to be 
degraded again.

The binding efficiency to gluthione sepahrose resin seemly depends on the 
protein expression and folding, which may be affected by some unknown factors, 
such as sonication?

I am going to try to express the protein with E.coli BL21 (DE3) LysS. so to 
avoid the sonication.

Someone may have such experience, and happy to share it.

Thanks!

Cheng

 

-- 
Cheng Luo,  Ph.D. 
BTK/Medicity Research Laboratory 
University of Turku
Tykistökatu 6 A
FIN-20520, Turku
Finland
	
Tel. +358 2 3338001
Fax  +358 2 3337000
Email  Cheng.Luo at utu.fi

URL:http://users.utu.fi/cheluo


Lainaus "Catherine L. Lawson" <lawson at rutchem.rutgers.edu>:

> Please try to resend your message to the xtallog bulletin board 
> (bionet-xtallography at net.bio.net).   Somehow there was no text!
> 
> Sincerely,
> 
> Cathy Lawson
> bionet.xtallography moderator
> 
> At 05:46 PM 11/18/2002 +0000, you wrote:
> 
> 
> 
> >---
> 
> 
> 
> 
> ***********************************************************************
> Catherine L. Lawson
> Laboratory for Structural Biology and Bioinformatics
> Department of Chemistry and Chemical Biology
> Rutgers University
> 610 Taylor Road, Piscataway, NJ 08854
> lawson at rutchem.rutgers.edu
> tel 732 445 8074
> fax 732 445 5312
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> http://roma.rutgers.edu/~lsbb
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