HELP: Coli is rearranging recombinant plasmid

Alexey Merz alexey at webcom.com
Thu Apr 23 08:38:39 EST 1998


Michael O'Donohue wrote:
> Alexey Merz wrote:
> > Right on. Make sure that the insert is cloned in an orientation
> > _opposite_ to the B-gal gene. Use a low copy vector. Maybe the DNA
> > seq. is inherently toxic but much more likely is that a poisonous
> > protein is getting made.
> >
> > Great vectors are the pWKS/pWSK vectors. These vectors are _identical_
> > to the pBluescript plasmids (same polylinker, F1 ori, etc) but have a
> > low copy origin and -better yet- WSK129 and WKS130 are Kanamycin
> > resistant instead of Amp resistant, meaning zero satellite colonies.
> > ref: Wang & Kushner, '91, _Gene_ 100:195-199.
         ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
> Great stuff! So where can one obtain this wonder vector?



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