P. aeruginosa transformation

Neil Saunders saunders at bio.vu.nl
Sat Feb 19 17:35:33 EST 2000


"Arnoud van Vliet" <avvliet at knoware.nl> wrote in message
news:950973761.877199 at news.knoware.nl...
> You will have to give more info, on sieze of plasmid, which selection etc.
> Anyway, best advice might be to try electroporation
> Arnoud
> From: Yong Jiang <yjiang at ucsd.edu>
> Newsgroups: bionet.molbio.methds-reagnts
> Sent: vrijdag 18 februari 2000 20:50
> Subject: P. aeruginosa transformation
> > Hi,
> > I am transforming P. aeruginosa with plasmid which can replicate in it
by
> > CaCl2 mediate method. Everytime I spread the no DNA transformed cells on
> > antibiotic plate, cells grow full of the plate. Could you give me amy
> > suggestion?
> > Best Regards!
> > Yong

I would also advise electroporation-there are a number of P. aeruginosa
protocols available.   But growth of non-transformed cells sounds like a
selection problem.  P. aeruginosa is resistant to a wide range of
antibiotics, what selection are you using?  For instance, ampicillin is no
good, you have to use carbenicillin at high concentration (250-500
microgram/ml).

Neil Saunders
--
Department of Molecular Cell Physiology,
Faculty of Biology,
Vrije Universiteit,
De Boelelaan 1087,
1081HV, Amsterdam,
The Netherlands

tel: +31 20 4447194
fax: +31 20 4447229

email: saunders at bio.vu.nl
http://www.crosswinds.net/~nfws/index.htm









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