Plasmid Isolation for over-expressed protein

Glenn Wall-Manning gmanning at wnmeds.ac.nz
Sun Sep 13 16:13:14 EST 1998


Sean Yoder wrote in message <35FA0BC5.9BBB401A at chuma.usf.edu>...
>    I'm having problems consistenltly isolating plasmid DNA from JM109
>host with a pCMV plasmid with a gene coding for a wall protein of S.
>mutans.  The plasmid seems to be over-expressing the gene and
>consequently lysing cells prematurely possibly due to the thin cell wall
>of E.coli.
Try growing your bugs in isotonic concentrations of sucrose.  This should
stop them popping prematurely.

--
Glenn Wall-Manning
Dental Research Group                 Standard disclaimers apply
Wellington School of Medicine
New Zealand






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