Independent transformants

Michael Lassner mikel51 at mother.com
Tue Nov 18 13:56:15 EST 1997


Several people asked me to post the responses to my question about
independent transformants.  In summary the answerers said that the
transformants are likely to be independent events.  It was also
suggested that the first batch of harvested seeds are unlikely to yield
transgenic plants, and that after a while the harvested seeds would no
longer yield transformants.


From:
           Maria Antonia LLuch Sanfeliu <lluch at farmacia.far.ub.es>
Every transformant from one infiltrated mature plant come from an
independent transformation event, so even in the first seed harvesting
you
obtain a pool of independent transformants. In conclusion these multiple
plants coming from the same mother plant and from the same infiltration
event are different from each other.


From:
             "Stefan Trentmann"
<trentsbl at mailhost.rz.ruhr-uni-bochum.de>
I usually wait for 3-4 weeks as well until I harvest seeds from
transgenic plants to ensure to get 1 up to 10 transgenics from each
infiltrated plant. I even discard the first couple of siliques
since they are not transformed anyway.
But to ensure for independant transformants I continue working
 with only one transgenic plant isolated from an  infiltrated
Arabidopsis
 plant. Nevertheless you also can receive independant events from one
infiltrated plant but this has to be prooven by Southern blot.

From:
           a-bent at uiuc.edu (Andrew Bent)
>From everything we have heard, the vast majority will be independent.


From:
           "LEVIN JOSHUA EXC CP US" <joshua.levin at usre.mhs.ciba.com>
i would be interested in seeing the replies to your msg.  i have
wondered the
same thing, but have been told that the later arising flowers would not
be
transformed at all.

From:
           "Tanja Singer" <tanja at snoopy.org.chemie.tu-muenchen.de>
I did a lot of vacuum infiltration on Arabidopsis plants and subsequent
Southern
anaIysis of the transgenes. I always harvested all the seeds of
infiltrated
plants at one time. My experience was that I never got the same agro
integration
pattern in two plants. My transformation rate is about 2 to 4
transformed plants
in 1000 screened seedlings.

From:
           j.g.turner at uea.ac.uk (j.g.turner)

My understanding of experiments reported on the network some years ago
is
that only buds at a certain juvenile stage during infiltration go on to
produce transgrenic seed. Apparently, buds which develop to that stage
at
some time after the infiltration do not usually produce transgenic seed.
This suggests that the proportion of independent transformation events
is
going to be the same, but the yield of transformants will depend on
which
harvest contains the siliques that were transformed. We get around 2%
transformants, which has been sufficient for our purposes. Good luck.
John






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