transformation

Andrew Bent abent at UX1.CSO.UIUC.EDU
Wed Mar 1 17:52:29 EST 1995


Colleagues -

I have fielded a number of similar questions recently about Agro strains
and Arabidopsis ecotypes that work well in the Bechtold et al.
Agrobacterium vacuum infiltration protocol for transformation of
Arabidopsis.  I have been able to reply based only on my own experience,
and it would clearly be helpful to many if we could gather a bit more
information from the community.

Replies will be summarized.  Any messages that are broadcast will be
stripped of names unless specifically requested (so that people feel a bit
more free to report failures and poorly replicated anecdotes as well as
successes).

The questions are:

1)  What Agro strains have worked well for you and your colleagues? Which
strains have not worked well?  Which ecotypes did you try?

2)  Which ecotypes have you successfully transformed?  Which haven't
worked?  Which ecotypes seem especially willing to be transformed?  Did the
Agro strain used seem to matter?

3)  Have you observed any primary transformants that are or are apparently
homozygous for the insert DNA?
      (This has a direct bearing on when the transformation event occurs.
For further discussion see:

Bechtold, N., J. Ellis and G. Pelletier, 1993.  In planta Agrobacterium
mediated gene transfer by infiltration of adult Arabidopsis thaliana
plants.  C. R. Acad. Sci. Paris, Life Sciences 316: 1194-1199.
     and:
Feldmann, K. A., 1992. "T-DNA insertion mutagenesis in Arabidopsis: Seed
infection/transformation." In: Methods in Arabidopsis Research. C. Koncz,
N. Chua and J. Schell, ed. World Scientific. London.)

4)  Do you have additional comments?


Thanks in advance  - PLEASE RESPOND!

Andrew Bent


-------------------------------------
Andrew Bent
Assistant Professor
Department of Agronomy
University of Illinois
Urbana, IL  61801

Phone 217-244-6308
Fax   217-333-4777





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