Plant Transformation and Regeneration

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Plant Transformation and Regeneration

• Gene Delivery Methods
• Agrobacterium tumefaciens-mediated T-DNA
  transfer
• Biolistic (gene gun)-mediated gene transfer
• Expression Cassette Assembly
• Critical components for transgene expression
• Vector Choices
• Tissue Culture-Based Gene Transfer
• Agrobacterium-mediated soybean transformation
• Biolistic-mediated maize transformation
• Transgene Analysis Integration patterns

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Crown-Galls Tumors tumefaciens
(Courtesy from Sharon von Broembsen)
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(Courtesy from Sharon von Broembsen)
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T-DNA Transfer From Agrobacterium To Plant Cell
T-DNA
Wild Type
RB
LB
Plant cell
Chromosome
Ti-plasmid
T-DNA
Disarmed
RB
LB
Chromosome
Ti-plasmid
Binary vector
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Biolistic PDS 1000/He
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Critical Components for Transgene Expression
MCS
RB
Binary vector
Expression Cassette
LB
T-DNA
GOI
Promoter
Terminator
Cloning vector
MCS
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Soybean Transformation and Regeneration Process
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Genomic Southern Blot of Transgenic Soybean
W-13-12
W-14-20
W-14-24
W-14-29
W-11-42
W-13-28
W-13-24
W-14-8
Kb 23.0 9.4 4.4
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Maize Transformation and Regeneration Process
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