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(AGROBACTERIUM
AGROBACTERIUM-
AGROBACTERIUM-MEDIATED
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Reading Materials
GENETICALLY MODIFIED
ORGANISMS (2003)
Yves tourtes
PLANT BIOTECHNOLOGY
(2003) - Adrian Slater, Nigel
Scott, Mark Fowler
Agrobacterium tumefaciens:
a natural tool for plant
transformation, Electronic
journal of biotechnology: Vol
1, No3, Dec 15, 1998
Essential requirements for plant genetic
engineering:
Agrobacteria
soil bacteria, gram-negative, related to Rhizobia
species:
tumefaciens - causes crown galls on many dicots
rubi - causes small galls on a few dicots
rhizogenes - hairy root disease
radiobacter - avirulent
Crown galls caused by
A. tumefaciens on
nightshade.
Gall on
stem
Gall on
leaf
Agrobacterium tumefaciens
cyt
AMP + isopentenylpyrophosphate isopentyl-AMP
(a cytokinin)
E. coli
Agrobacterium
The intermediate
vector that contains the Agrobacterium
foreign DNA is
introduced by
conjugation into
Agrobacterium that
contains the co-
integrative vector.
E. coli
The intermediate vector is not
stable in Agrobacterium.
Homologous recombination
between the intermediate vector HOMOLOGOUS
and the cointegrative vector RECOMBINATION
results in transferring the foreign
DNA to the cointegrative vector.
These vectors are designed so that once the foreign DNA, is
integrated into the cointegrative vector, it is located between its border
sequences.
2 Common Transformation Protocols
1. Leaf-disc transformation - after selection and
regeneration with tissue culture, get plants with
the introduced gene in every cell