temp. sensitive plasmid

Jim Miller jrm at lilly.com
Tue Oct 15 04:18:04 EST 1996


Jeff Huckaby wrote:
> 
> Netters
>         I need a bacterial plasmid that has a ts origin of replication or some
> other mechanism such that when placed at the non-permissive temp. it no
  ^^^^^^^^^^^^^^^   
> longer remains in the cell. 

Snip

Any Help Would be Appreciated.

Track down a PolI mutant of E. coli.  If my memory serves me right, colE1-based 
plasmids require PolI for replication and the ts mutants of PolI (dnaA-?), when raised 
to the none-permissive temperature,  will "cure" the cell of the plasmid.  BTW, all 
pBR322 (aka pUC) vectors are derived from colE1-am I right?

Hope this helps.

Jim Miller
Lilly Research Labs
jrm at lilly.com

> Thanks
> Jeff
> 
> ==============================================================================
> Jeff Huckaby                                                      Jeff Huckaby
> Department of Biology                                                  426 OML
> email huckabyj at minerva.cis.yale.edu                            Yale University
> office:(203) 432-3877                                      New Haven, CT 06511
> WWW http://www.yale.edu/~huckabyj/topbio.html               Lab:(203) 432-3982
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