Plant Transformation and Regeneration - PowerPoint PPT Presentation

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Plant Transformation and Regeneration

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Plant Transformation and Regeneration – PowerPoint PPT presentation

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Title: Plant Transformation and Regeneration


1
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

2
Crown-Galls Tumors tumefaciens
(Courtesy from Sharon von Broembsen)
3
(Courtesy from Sharon von Broembsen)
4
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
5
Biolistic PDS 1000/He
6
Critical Components for Transgene Expression
MCS
RB
Binary vector
Expression Cassette
LB
T-DNA
GOI
Promoter
Terminator
Cloning vector
MCS
7
Soybean Transformation and Regeneration Process
8
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
9
Maize Transformation and Regeneration Process
10
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