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Integration into the URA3 locus

Amine Noueiry anoueiry at facstaff.wisc.edu
Mon Jun 24 15:09:03 EST 1996


Hi everybody

	I am trying to integrate a construct into the URA3 locus. I have two 
constructs containing a partial deletion of that gene. The constructs 
have either LEU or HIS selection. Upon cutting with a restriction enzyme 
(the sites are different for the two plasmids) in the URA3 locus and 
transformation into my URA3+ HIS- LEU- strain and selecting on LEU or 
HIS plates, I get no colonies. As a control, I am using the same LEU 
integration vector with a different locus cloned into it to integrate, 
and this one gives me many (100s) colonies. Nobody I know has ever tried 
integrating into the URA3 locus. If you do and you read this, please 
email me with advice. I would greatly appreciate it. Thank you in 
advance, 

						Amine





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