Expression library screening with fusion proteins

Petri T. Kursula pkursula at zombie.oulu.fi
Tue Aug 15 07:36:59 EST 1995


I am starting to look for proteins that bind a specific protein domain 
that we are producing as a GST fusion protein. I wonder if anyone has any 
experience in screening an expression library with such a fusion protein, 
using anti-GST anibody and a secondary antibody coupled with alkaline 
phosphatase to visualize the positive clones, i.e. the ones that bind the 
fusion protein. 

What kind of conditions should one use to make the fusion 
protein bind the ones on the filter made from the expression library 
plates? Also, how easy would you think it would be to totally wash such a 
filter so you could use a different fusion protein to screen the same 
filter? What kind of conditions should you absolutely NOT use? Any help 
will certainly be appreciated!


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|					|    	       /      \	
|	Petri Kursula	 		|	       | -  o |
|       Department Of Pathology		|	       |  ..  |	
|	University Of Oulu		|	       | \__/ |	
|					|		\____/
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