Nuts and Bolts of Genetically Engineered Crops

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Principles of Applied Biotechnology

• AGENDA OUTLINE

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Transgenic Crops

• Do you know your GMOs?
• Do you know your Events?
• Is it really a non-GMO?

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Non-GMO vs. GMO
Change a gene
Insert a new gene
Make New Protein
Alter Protein
Change Plant or Seed traits
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Pop Quiz
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Non-GMO
Seed
Plant
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Non-GMO
Cell
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Non-GMO
Cell
DNA
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Non-GMO
Cell
Gene
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Non-GMO
Cell
Protein
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Non-GMO
Cell
Trait!!
Protein
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Non-GMO
SeedTrait
Cell
Plant Trait
Trait!!
Protein
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Non-GMO
SeedTrait
Cell
Plant Trait
Mutation
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Non-GMO
SeedTrait
Cell
Plant Trait
Mutation
Alter Protein
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Non-GMO
SeedTrait
Cell
Input Trait Herbicide Resistance
Mutation
New Trait!!
Alter Protein
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Non-GMO
Output Trait Oil or Protein
Cell
Input Trait Herbicide Resistance
Mutation
New Trait!!
Alter Protein
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GMO
Cell
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GMO
Cell
GMO Genetically Engineered
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GMO
Cell
New gene
Gene from any organism
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GMO
Cell
New gene
Insert !!
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GMO
Cell
New gene
Chromosome breaks
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GMO
Cell
New gene Transgene
Transgene part of chromosome
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GMO
Cell
Transgene
Make New Protein
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GMO
Cell
Make New Protein
New Trait!!
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GMO
Output Trait Oil or Protein
Cell
Input Trait Herbicide Resistance
Make New Protein
New Trait!!
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GMO
Input Trait They are the same as non-GMO
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GMOs
Output Trait They are different than on-GMO
Input Trait They are the same as non-GMO
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Non-GMO vs. GMO
Change a gene
Insert a new gene
Make New Protein
Alter Protein
Change Plant or Seed traits
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Crop Genetic Engineering Process
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Crop Genetic Engineering Process
Bacillus thuringiensis makes BT protein toxic
to ECB
1. Discover organism with a trait you want in
your crop plant
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Crop Genetic Engineering Process
Bacillus thuringiensis BT
2 and 3. Extract the DNA and clone the gene that
encodes the desired protein
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Crop Genetic Engineering Process
4. Modify the gene
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Crop Genetic Engineering Process
5. Transform cultured corn cells
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Crop Genetic Engineering Process
6. Clone a plant from the cell
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Crop Genetic Engineering Process
7. Breed elite lines to the trangenic plants
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Process is similar RoundupReady
Agrobacterium
Soybean cell
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Molecular cellular Biology
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FLASH Animation
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DNA extraction PPT
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How to Clone a Gene
Cloned Gene A copy of a gene.
Step 1 Obtain the organism.
Step 2 Isolate the DNA.
Gene of interest (Bt toxin gene)
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Step 3 Cut into pieces with restriction
enzymes.
Step 4 Cut the plasmids with the same a
restriction enzyme.
Plasmids Small circles of DNA that contain a few
genes. Naturally found in bacteria.
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Step 5 Add the two cut up samples of DNA into
the same tube, add ligase enzyme which will
combine the ends of cut DNA.
Ligase enzyme
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Step 6 Introduce plasmids into living E. coli
bacteria cells
E. coli bacteria cell that does not have a
plasmid
Transformed E. coli bacteria cell that has a
plasmid
E.coli Bacteria has its own chromosome of DNA
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Transformation of Bacteria
Step 6 (continued) Transformation of bacteria,
some bacteria get a plasmid with an inserted
gene, some do not.
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Step 7 Bacteria are plated on media and grow
into colonies.
Antibiotic resistance gene OK
Antibiotic media plates
Insert in Blue Gene
Blue Gene OK
Antibiotic resistance gene OK
Bacteria that do not get a plasmid will not grow
on antibiotic media
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Step 8 Select the white bacterial colonies.
You do not know which colony has the gene you
are interested in.
Gene Library A collection of cloned genes from a
particular organism.
Step 9 Repeat to make a large collection.
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Step 10 Screen the library.
Grow the bacteria with the gene of interest to
make many copies of it.
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Gene Expression
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nucleus
cytoplasm
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DNA
chloroplast
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DNA Genes
chloroplast sugars
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DNA Genes
chloroplast amino acids
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chloroplast
nucleus
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DNA (genes)
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Gene is turned on
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Make a message (RNA)
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Make a message (RNA)
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Make a message (RNA)
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Make a message (RNA)
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Make a message (RNA)
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Message (RNA) leaves nucleus
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Message is read to make a protein (enzyme)
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Message is read to make a protein (enzyme)
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Message is read to make a protein (enzyme)
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Protein goes to work in the cell
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Protein goes to work in the cell