Which Agrobacterium strain?
Leonard N. Bloksberg
bloksber at pilot.msu.edu
Mon Jan 17 16:09:00 EST 1994
In Article <1994Jan17.130735.1 at molbiol.ox.ac.uk> "mknight at molbiol.ox.ac.uk" says:
> Can anyone out there enlighten me as to what goes into the choice of which
> Agrobacterium strain to use for transformations? I'm recommended by most people
> to use C58 and told that LBA4404 is a bad idea. Why is this?
> Many thanks!
> Marc Knight
> ==== ======
LBA4404 is not really a bad idea, it's just not the best. When people have
bothered to do side by side controled comparisons, Nopaline strains (eg,
C58) give higher rates of transformation with transgenes. Also, some of the
Octapine strains (eg the parent of LBA4404) still retain some of their onc
genes, and can yield strange and mostly sterile transgenics (not so for
LBA4404, as far as I know). LBA4404 contains the plasmid pAL4404 which is
a HindIII partial digest of the original ti plasmid, and now has only the
vir region left. My favorite is LBA4301 with the plasmid pUCD2614. The
strain LBA4301 has the advantage of being rec-. It was originally an
Octapine strain, but was cured of it's plasmid. The plasmid pUCD2614 was
made by cloning NotI fragments of the C58 Ti plasmid into pUCD1002 to
reconstruct the nopaline vir region only. I believe you can find the
reference in Plasmid, by Rogowski and Kado, from around 1988 (I think).
This combo (LBA4301 and pUCD2614) form a very efficient host and vir region
that can transfer any tDNA with high efficiency, and it's rec- to boot. I
recomend you write directly to Dr. C. I. Kado to get this strain. Good Luck.
. Dr. C. I. Kado
. Dept. of Plant Pathology
. University of California, Davis
. Davis, CA 95616
. Tell him I sent you :-)
. Leonard N. Bloksberg
. Dept. of Crop and Soil Science
. Michigan State University
. East Lansing, MI 48824
. bloksber at pilot.msu.edu
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