dicistronic plasmid - help

Jillian Johnston johnstoj at acs2.acs.ucalgary.ca
Fri Jul 31 10:58:25 EST 1998


I have been trying unsuccessfully to make a plasmid that will
express both GFP as a marker and another gene.  Both genes are
driven by identical CMV promoters and terminated with identical
SV40 polyA.  I know that repeated sequences are difficult to
clone so I am trying to have the genes going in opposite
directions.  The plasmid I've been using is pBluesciptSK
(Stratagene) and DH5a for transformations. I get no colonies but
the ligation reaction is working, as seen on a gel.  Without
going into all the details, which I am certainly willing to 
provide, I was wondering if anyone has any insights into a
solution.
Thanks

--
Jillian Johnston
Dept. of Biological Sciences       Guider
Univ. of Calgary         	   30 Pathfinders, Tanisi District




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