Since I wrote about a Tobacco leaf infiltration protocol that works in
Arabidopsis to check GFP fusion constructs, I have been inundated with
requests for the protocol so I will post it here. I don't think it is
substantially different to the one from the John Innes Centre. A couple
of qualifying statements:
1) It works, just not as well as in Nicotiana.
2) The trick seems to be to get the leaves at just the right stage. You
want nice healthy, juicy plants from which you infiltrate into rosette
leaves that are still expanding. The protocol really will not work on
old leaves even in tobacco.
3) If you are just wanting to check constructs for protein localization,
grow some tobacco and use the protocol. It works so well it is very
satisfying. From the completion of vector construction, it takes less
than a week to grow the Agros, infiltrate the leaves and see expression.
The tobacco plants can be young and small and don't require a huge
amount of space or time to maintain. Once you know that your construct
works you can either, i) try the protocol on Arabidopsis or ii) use
floral dip to make stable Arabidopsis lines
The protocol is attached - good luck! And please provide feedback to
the list if you invent a way to get better expression in Arabidopsis.
C. John Runions, Ph.D.
School of Biological and Molecular Sciences
Oxford Brookes University
email: jrunions at brookes.ac.uk
phone: +44 (0) 1865 483 964
-------------- next part --------------
A non-text attachment was scrubbed...
Name: Tobacco leaf infiltration.pdf
Size: 24267 bytes
Desc: not available
Url : http://www.bio.net/bionet/mm/arab-gen/attachments/20051209/6587776f/Tobaccoleafinfiltration-0001.pdf