His-tags and DNA-binding

Matt Thomas mthomas at hsc.usf.edu
Fri Jan 12 14:54:15 EST 2001


My old lab used to make his tagged TBP and it bound TATA boxes just fine.
It will probably be a case specfic issue and you'll just have to try it.  Or
put on one of those leaders with a protease site and cut off the tag to do
some controls.
-- 
___________________________________________
Matthew Thomas, Ph. D
H. Lee Moffitt Cancer Center, University of South Florida
mthomas at hsc.usf.edu

"I refrain from publishing for fear that disputes and
controversies may be raised against me by ignoramuses."
   Sir Isaac Newton, correspondence to Liebniz

> From: Matt Hicks <matth at biols.susx.ac.uk>
> Organization: University of Sussex
> Newsgroups: bionet.molbio.proteins
> Date: Thu, 11 Jan 2001 10:51:29 +0000
> Subject: His-tags and DNA-binding
> 
> Hi Folks,
> Has anyone come across a situation where a 6XHis tag has affected the
> DNA-binding of a DNA-binding protein? (the pH is 8 so unlikely to be +ve
> charge). I know the folding could be affected. Just wondered if anyone
> had seen such a phenomenon...
> Thanks,
> 
> **************************************************
> 
> Dr. Matt Hicks
> School of Biological Sciences
> University of Sussex
> Falmer
> BN1 9QG
> UK      
> 
> Tel:   +44(0)1273 678923
> Fax:   +44(0)1273 678433
> email: matth at biols.sussex.ac.uk
> 
> **************************************************
> 






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