Hi netters, I have a question:
I am working with a Pseudomonas sp. that contains a target gene
(an halidohydrolase) that I am currently studying.
Such gene could be carried on a mobile genetic element, as it
has been previously reported in similar circunstance, and
as its expression behaviour lets me to believe.
In order to test such evenience I would like to transfer in such
Pseudomonas a plasmid of the RP4 family (RP4.5) and see if the
target gene (chromosomically located) recombines with
the plasmid. Later, I want to select some transconjugants as
donors and mate them with P. putida KT2440, and check if the
plasmid-determined antibiotic resistance markers are
cotransferred with the phenotype carryied on the mobile
element.
Unfortunately I don't know were to purchase RP4 family
plasmids or other plasmids of similar properties. Moreover,
I don't know if such plasmids are the best choice.
If someone on the net could inform me on such topic
I would be very grateful.
Tanks a lot,
-- Vinz
Vincenzo Nardi-Dei
Laboratory of Microbia Biochemistry
Institute for Chemical Research
Kyoto University
Uji-Campus 611 Japan
