[Arabidopsis] Tobacco leaf infiltration protocol

John Runions jrunions at brookes.ac.uk
Fri Dec 9 13:42:39 EST 2005



Hi All,

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
Oxford, UK
OX3 0BP

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
Type: application/pdf
Size: 24267 bytes
Desc: not available
Url : http://www.bio.net/bionet/mm/arab-gen/attachments/20051209/6587776f/Tobaccoleafinfiltration-0001.pdf


More information about the Arab-gen mailing list