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more q on pseudomonas vectors

Don Chen chend at ucs.orst.edu
Mon Dec 18 10:13:04 EST 1995


Hi:

I recently asked for information (in the methods newsgroup) about
vectors which either work in pseudomonads alone or in them and E.coli.
I received good information from Mark Strom
<strom at listeria.nwfsc.noaa.gov> about the pMMB66 series of vectors as
well as types of expression and shuttle vectors.  I am aware of the
Tn5 cassettes which are used to randomly (I think) mutagenize the
genomic DNA.  Tim Barnett <t.barnett at path.utas.edu.au> pointed out
both pUCP19 and the recently described pBBR1MCS.  It is the latter
which interests me now.  I tried to send email to
<petersonk at biotechnet.com> but I got back a note that it was bounced. 

I would like to get a hold of the vector since it appears to be able
to replicate in both E. coli and Pseudomonas (fluorescens is the
species I would like to use).  As I understand it, pBBR1MCS can be
transformed into both cells by electroporation.  Mark has given me a
recipe which he used in Ps. aeruginosa.  Is also possible to transform
those cells by CaCl2 precipitation? or is electroporation far superior
(any references)?

Beyond this, I am curious if there is a good book or review on
Pseudomonas and characteristics of the various species?  The library
doesn't seem to have any?

In that vein, Fred Hyde <hydex004 at gold.tc.umn.edu> has pointed out
that the MIDI method of characterizing bacteria based on fatty acid
methyl esters.  Has anyone who has used this method any comments on
its utility?

Thanks for any help you have given, and for future aid.

Don

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