pGreen discussion group

[Jonathan Rees]

Dear Collegues,

This announcement is aimed at people involved in Plant Transformation 
- please disregard otherwise.

pGreen is a small (<5kb) binary PLANT transformation and expression 
vector that allows users to routinely subclone without the problems 
of the larger pBin series by virtue of a 10kb helper plasmid (pSoup) 
which carries several of the required functions.  Please see the 
reference below:

Roger P. Hellens, E. Anne Edwards, Nicola R. Leyland, Samantha Bean 
and Philip M. Mullineaux (2000) "pGreen: a versatile and flexible 
binary Ti vector for Agrobacterium-mediated plant transformation", 
Plant Mol. Bio. 42: 819-832.
Also see http://www.pgreen.ac.uk/ for more details of the plasmid.

It has come to my attention and has been my experience that while 
high transformation efficiencies may be routine with pBin series 
vectors using the simple floral dipping method, a number of users 
have not experienced the same or consistent success with the pGreen 
system.  I have set up a Bulletin Board specifically to discuss the 
use of pGreen, problems with pGreen, and improvements to protocols 
involving handling of pGreen especially with regard to Arabidopsis 
transformation.  The board is freely available at:

http://pluto.beseen.com/boardroom/n/58644/

In short, the aim of the bulletin board is to come up with a 
'never-fail' optimized protocol to allow the routine use of this 
plasmid and its increasing number of derivatives with first-time 
success.

I have started off the proceedings with a comment about the safe use 
of the tetracycline resistance gene in Agrobacterium, and how NOT to 
use it with Agrobacterium C58 derived strains.

I hope to collate everyone's input and repost a link to a summary of
what people thought.  {The board is hosted on a free-site, so
apologies in advance for the nasty adverts!  It has been set up in
the absence of any equivalent discussion group}


best regards,

Dr Jonathan Rees

PS. I am a pGreen user, or 'pGreener', and I have nothing whatsoever 
to do with the plasmid's development!