genetic engineering. what is genetic engineering? definition: process of changing an organism’s...
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Genetic Engineering
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What is genetic engineering?
Definition: process of changing an organism’s genetic material to produce a new and useful result
This results in a transgenic organism (organism containing recombinant DNA)
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Tobacco Plant that has had a firefly gene inserted into its cells, so it “glows”.
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Why make transgenic plants and animals?
Food production Make plants that are resistant to pesticides and/or diseases
(Round-up® ready crops, crops resistant to certain viruses) Engineer food to stay fresh longer and ripen at the store Engineer some animals to grow larger without using
antibiotics or growth hormone. Protein production
Cows and goats could produce human proteins in milk Even plants can be used to produce human proteins Vitamin A in “golden rice”
Medical Research Use the “glow” gene to follow the expression of another
gene. Edible vaccines.
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How does genetic engineering work? The desired (“donor”) gene is
isolated from the organism’s DNA using a restriction enzyme
Bacterial plasmid (“cloning vector”) is cut with same restriction enzyme Sticky ends of desired gene and plasmid
should be complementary Ligase (an enzyme) is added to help
seal sticky ends of plasmid and desired gene together
Last step: put the recombinant plasmid back into the bacteria
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Example: Insulin Production
Protein that codes for insulin production is cut from human DNA using restriction enzyme (EcoRI)
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Example: Insulin Production
Plasmid (from E.coli) is also cut with EcoRI Sticky ends are now
exposed on both human DNA and plasmid DNA
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Example: Insulin Production
Ligase seals sticky ends of desired DNA and plasmid DNA
Recombinant DNA is inserted into a bacterium
Bacterium divides, producing 100’s of bacteria with the desired gene.
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Why insert recombinant DNA into a bacterium?
Bacteria reproduce rapidly (some every 20 minutes) Inserted gene (e.g. insulin) is copied
(cloned) whenever bacteria make a copy of the plasmid
Results in a large amount of desired protein (insulin) in just a few days
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How can you insert recombinant DNA into a host cell?
Transformation: use electricity, heat, or CaCl2 to “shock” cell into taking up plasmid
Microinjection: inject plasmid DNA directly into cell using microneedle
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How can you insert recombinant DNA into a host cell?
Gene gun: coat tiny gold pellets with recombinant plasmid DNA and “shoot” into cell
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How can you insert recombinant DNA into a host cell?
Viral vectors: Desired gene is placed into viral DNA and used to “infect” an organism’s cells without causing disease (virulence gene removed)