Looking for a vector with a Kill function
alexey at webcom.com
Wed Apr 22 20:22:36 EST 1998
Craig Sherburne wrote:
> I am searching for a vector able to replicate in E. coli, that contains an
> inducible kill function, for use in recombination experiments (plugging a disrupted
> gene back into the wild type). If anyone knows of a commercially available vector,
> or a Lab with such a construct I would appreciate the information.
> Craig Sherburne
> csherbur at gpu.srv.ualberta.ca
How about the F plasmid ccdB gene, which poisons DNA gyrase? In
the presence of ccdA it's nontoxic, but in the absence of ccdA
it's quite lethal. The F plasmid seems to use this system to
murder segregants not carrying F, because CcdB protein has a
longer half life than CcdA. Shut off synthesis of both and the
cell dies. A Medline search should yield some refs.
Alternatively, it is possible to counterselect against tet resistance;
I think that this is described in Jeffery Miller's book.
There are lots of other approaches, of course.
Alexey Merz | URL: http://www.webcom.com/alexey | email: alexey at webcom.com
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