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Plant Transformation Method (Physical (Particle bombardment (Can transform…
Plant Transformation Method
Physical
Particle bombardment
Can transform organelles
Expression vector is precipitated outo tungsten or gold
Can use on cells with cell walls
Transient expression
Microprojectile
Mutiple copies of transgene insert
Electroporation
Biolistics ( gene gun)
Microinjection
Silica / carbon filters
Whiskers-mediated transformation
target cells, fibers and plasmid DNA is placed in a tube and mixed by vortex.
treated corn tissue is placed on selection media of the regeneration of GM seedlings
the use of silicon carbide
functions as needles that enabling the entry of DNA into cells during mixing process
Chemical
Polyethylene glycerol (PEG)-mediated DNA transfer
oldest reliable method for plant transformation
protoplasts are treated with DNA in the presence of PEG and Ca++
Agrobacterium Ti was introduced into petunia protoplast
Calcium phosphate
In planta
Virus-mediated
Agrobacterium mediated gene transfer
Agrobacterium tumefaciens
(Ti plasmid)
Ti plasmid
Origin of replication
Conjugation genes
Vir region
24 vir genes
8 operons
T DNA region
Containing genes that encode for synthesize of hormones and opines
25-base-pair repeats
initiated at right border and terminated at left border
24 kb
Opine catabolism
Agrobacterium rhizogenes
(hairy roots)
Agrobacterium radiobacter
(avirulent)
Biological
Conjugation
Transduction