In article <rfriedma-ya023180001110961229230001 at netnews.upenn.edu>,
rfriedma at thunder.ocis.temple.edu (Richard Friedman) wrote:
> In article <Pine.LNX.3.91.961009171722.8186A-100000 at morpheus.wustl.edu>,
>gerry at MORPHEUS.WUSTL.EDU (Gerald Downes) wrote:
>> > Netters!
> > I received a pamplet from Clontech that describes their
> > pZErO cloning vector. Of course it *sounds* like heaven on earth (ease
> > of use with no background) but is there a catch? Does anyone have
> > experience with this vector or another that is supposed to give
> > virtually no backgroud (CIP treated vectors excluded).
> > Gerry
>> The catch is that the plasmid can only be grown in a gyrA44 (I think)
> mutant. I don't think the standard gyrA mutants (such as DH5alpha) will do
> the job. As for the stain used in selection, probably any gyrA+ strain will
Can't you bulk up these plasmids in lacIq strains where expression from
Plac should be turned off and the ccdB protein wont be made or is it too
potent at even these low levels? What about F' strains - wont they contain
the ccdA antidote and allow growth of the original vector? XL1 Blues are
both F' and lacIq, has anyone tried growing up pZero in these?
John Dixon Lab 44 (1223) 334131
Wellcome/CRC Institute Fax 44 (1223) 334134
Department of Genetics
United Kingdom e-m: jpcd0 at mole.bio.cam.ac.uk