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Application of PCR

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Generation of Transgenic Plants leaf single cell transformation of cell (gene transfer) protoplast transformation gene gun Agrobacterium tumefaciens – PowerPoint PPT presentation

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Title: Application of PCR


1
Generation of Transgenic Plants
leaf
single cell
transformation of cell (gene transfer)
protoplast transformation
gene gun
Agrobacterium tumefaciens
regeneration
transformed cell
callus culture
transgenic plant
2
Cultivation of Transgenic Plants in 2005
3
Gene Transfer by Gene Gun (Biolistic)
4
Gene Transfer by Gene Gun in Plant Cells
pressure compartment
suppport for DNA-coated particles
breaking plate
DNA-coated particles
scattering grid
plant tissue in petri dish
5
Generation of Transgenic Plants
leaf
single cell
transformation of cell (gene transfer)
protoplast transformation
gene gun
Agrobacterium tumefaciens
regeneration
transformed cell
callus culture
transgenic plant
6
Generation of Transgenic Plants
leaf
single cell
transformation of cell (gene transfer)
protoplast transformation
gene gun
Agrobacterium tumefaciens
regeneration
transformed cell
callus culture
transgenic plant
7
RoundupReady Soy Beans (RRS)
  • Soy beans are processed to soy oil and bruised
    grain
  • 30.000 food products contain soy oil (chips, ice
    cream, bakery products, candy bars...)
  • RoundupReady soy beans are resistant against the
    herbicide Roundup, glyphosate,
    N-(phosphonomethyl)-glycine) sold by Monsanto
  • Glyphosate inhibits the enzyme EPSPS
    (3-enolpyruvyl-shikimate-5-phosphate-synthase)
    which is needed for the biosynthesis of aromatic
    amino acids
  • Cells die without aromatic amino acids

8
Glyphosate - Mode of Action -
9
RoundupReady Soy Beans (RRS)
  • Soy beans are processed to soy oil and bruised
    grain
  • 30.000 food products contain soy oil (chips, ice
    cream, bakery products, candy bars...)
  • RoundupReady soy beans are resistant against the
    herbicide Roundup, glyphosate,
    N-(phosphonomethyl)-glycine) sold by Monsanto
  • Glyphosate inhibits the enzyme EPSPS
    (3-enolpyruvyl-shikimate-5-phosphate-synthase)
    which is needed for the biosynthesis of aromatic
    amino acids
  • Cells die without aromatic amino acids
  • RoundupReady soy beans contain a
    glyphosate-resistant bacterial gene from
    Agrobacterium tumefaciens CP4
  • 30 reduction of use of glyphosate
  • RoundupReady soy beans is sold by Monsanto
  • USA produces 60 million tons of soy beans per
    year (50 of worldwide harvest)
  • ---------------------

10
Vector for Glyphosate Resistance
11
T-DNA from Agrobacterium tumefaciens
gene of interest
plant promoter
plant resistance gene for selection
terminator
left border of T-DNA
right border of T-DNA
bacterial resistance gene for selection
ori /origin of replication
bacterial promoter
12
Labeling
  • Food, food ingredients and food supplement have
    to be labeled when they are GMO, consist of GMO
    or contain material of GMO
  • Labeling is required when the percentage of GMO
    material is greater than 0.9 in food containing
    a mixture of normal and GMO material
  • Examples
  • Food or food supplement are GMOs GM tomato, GM
    patato, GM corn, GM fish...
  • Food or food supplement contain GMOs GM bacteria
    in yogurt, GM yeast in beer...
  • Food or food ingredients are derived from GMO
    oil from GM soy beans or GM corn, sugar form GM
    sugar beet, starch from GM corn...
  • Food or food supplement are isolated from GMO
    Vitamin C, vitamin B2, glutamate...
  • Labeling is not required when
  • meat, eggs or milk comes from animals fed with
    GMO plants
  • Material from GMO fed to microorganisms, e.g.
    yeast cells fed with starch from GM corn
  • enzymes isolated from GMO chymosin produced by
    GM bacteria
  • ---------------------------------------------
  • GMO genetically modified organism

13
End
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