GST-fusion protein

Cheng Luo cheng.luo at utu.fi
Mon Nov 18 14:15:12 EST 2002



> 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
> > http://rutchem.rutgers.edu/~lawson
> > http://roma.rutgers.edu/~lsbb
> > *********************************************************************
> > 
> > 
> 
---



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