Clontech library problem

Seng-Lai Tan sltan at u.washington.edu
Fri May 9 14:27:35 EST 1997


i think clontech sells antibodies against the GAL4 BD and AD domains so
you can test expression of your hybrid proteins.  I have also used
the yeast 2-hybrid system to pull out proteins that make a lot of sense
but they were all out of frame wrpt the vector sequence (pACT2).  My guess
is that somehow the plasmid sequences were mutated when i passed
transferred them into E. coli (i used DH5a) for amplification and
purification. 

On Fri, 2 May 1997, Jennifer Glick wrote:

> 
> Hello netters!
> 
> I have a Clontech two-hybrid (Matchmaker) library which is giving me 
> fits.  I have pulled out four clones which all make sense.  However, 
> three of the four are out of frame w/r/t the vector sequence from 
> Clontech.  While the clones are all proteins which have been identified, 
> there are no antibodies for me to test if they are indeed what is being 
> expressed.  My question is, has anyone out there used this library???  
> Its a human brain library in pACT2.
> 
> thanks,
> Jennifer Glick                                jbelyea at acpub.duke.edu
> Duke University Medical Center			(919) 613-8615 Tel.
> Durham NC 27707					(919) 613-8642 Fax.
> 




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