Floral Transformation - patent granted to Paradigm Genetics Inc.
The invention disclosed in this patent comprises an alternative protocol to
adjust the cell density of the Agrobacteria used in the transformation
step. This method should be useful in high throughput transformation protocols.
According to the INPADOC patent family information, this patent is granted only
in the United States.
Specific patent information
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
6353155 B1
- Earliest priority - 30 June 2000
- Filed - 30 June 2000
- Granted - 5 March 2002
- Expected expiry - 30 June 2020
|
Title - Method for transforming plants
Claim 1
A method for preparing a transgenic plant or seed comprising:
a) growing a suspension of Agrobacterium cells until the optical
density of the suspension is about 2 to about 2.4 at a wavelength of 600
nanometers, wherein said Agrobacterium cells contain at least one
plasmid having a DNA sequence of interest flanked by T-DNA borders;
b)
diluting said Agrobacterium cells of said suspension with an aqueous
medium so that the optical density of the suspension is reduced to about 0.6 to
about 1.5, wherein said diluting is not followed by pelleting via centrifugation
with subsequent resuspension; and
c) treating the flower of said plant with
said diluted suspension so that the Agrobacterium cells in said diluted
suspension can transform said plant with the DNA-sequence of interest;
d)
optionally, cultivating said treated plant to produce seed; and
e)
optionally, growing plants from said seed and selecting for transgenic plants
having said DNA sequence of interest.
|
Claim 10
A method for preparing a transgenic plant or seed comprising:
a) growing a suspension of Agrobacterium cells until growth of
Agrobacterium cells in the suspension is substantially completed,
wherein said Agrobacterium cells contain at least one plasmid having a
DNA sequence of interest flanked by T-DNA borders;
b) diluting said
suspension with an aqueous medium to reduce the concentration of
Agrobacterium cells and any other components in the growth medium and
allow the Agrobacterium cells to infect the plant without harming it,
wherein said diluting is not followed by pelleting via centrifugation with
subsequent resuspension;
c) treating the flower of said plant with said
diluted suspension so that the Agrobacterium cells in said diluted
suspension can transform said plant with the DNA-sequence of interest;
d)
optionally, cultivating said treated plant to produce seed; and
e)
optionally, growing plants from said seed and selecting for transgenic plants
having said DNA sequence of interest.
|
Claim 17
A method for preparing a transgenic plant or seed comprising:
a) growing a suspension of Agrobacterium cells until growth of
Agrobacterium cells in the suspension is substantially completed,
wherein said Agrobacterium cells contains at least one plasmid having a
DNA sequence of interest flanked by T-DNA borders;
b) diluting said
Agrobacterium cells in said suspension with about 2 to about 10 volumes
aqueous medium per volume of suspension, wherein said diluting is not followed
by pelleting via centrifugation with subsequent resuspension;
c) treating
the flower of said plant with said diluted suspension so that the
Agrobacterium cells in said diluted suspension can transform said plant
with the DNA-sequence of interest;
d) optionally, cultivating said treated
plant to produce seed; and
e) optionally, growing plants from said seed and
selecting for transgenic plants having said DNA sequence of interest.
|
United States patent US 6353155 claims
- a method to transform floral tissue in vivo by treatment with an
Agrobacterium cell suspension adjusted to a certain density by dilution
with an aqueous solution.
- The claims exclude pelleting by centrifugation after the dilution step, but
not by other means.
- Claim 10 states that Agrobacterium should be allowed to infect
a plant without harming it. The patent does not define what "harm" means.
The invention comprises an alternative protocol to adjust the cell density of
the Agrobacteria used in the transformation step. According to the inventors,
centrifugation and resuspension are avoided by direct dilution of the bacteria
in an aqueous solution, thereby allowing sequential treatment of many more
plants. The method is well-suited for the transformation of Arabidopsis
thaliana (Brassicaceae), a model plant where such a high throughput
transformation protocol might be required. No other plants are used in the only
example provided as part of the patent specification.
|
Paradigm Genetics Inc.
|
Note: Patent information on this page was last updated on 23 February 2006.
The information contained in this page was believed to be correct at the
time it was collated. New patents and patent applications, altered
status of patents, and case law may have resulted in changes in the
landscape. CAMBIA makes no warranty that it is correct or up to date at
this time and accepts no liability for any use that might be made of it.
Corrections or updates to the information are welcome. Please send an email to
info@bios.net.
There are no comments.