Looking for a vector with a Kill function

Alexey Merz 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.

Good luck, 

-Alexey
..........................................................................
Alexey Merz | URL: http://www.webcom.com/alexey | email: alexey at webcom.com
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