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Discover the world of biotechnology and its various applications, from genetic engineering to the production of recombinant DNA. Explore the benefits of biotechnology in medicine, agriculture, and industry.
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What Is Biotechnology? • Any technique that uses living organisms or substances from those organisms to make or modify • a product • improve plants or animals • to develop microorganisms for specific uses
WHAT COULD SOME POSSIBLE REASONS BE???? • Adding a clotting agent to the blood that would help dissolve clots. • TPA--- DONE!!!!! • What if: • Cancer free genetics • HIV free genetics • Creating something that was resistant to these diseases???IS IT POSSIBLE?
Genetic Engineering • Genetic engineering is taking one or more genes from one organism and ….. • Transferring them to another organism • Putting them back into the original organism in different combinations GMO’s: • Transgenic Organisms: Bt Corn ( pest resistant) • Recombinant DNA (rDNA): rInsulin
What are the Benefits of Biotechnology? • Medicine • Human • TPA, Dissolve blood clots • Cancer Treatments • Embryo Screening • Veterinary • Hybrid Dogs • Surrogate Cows • Environment • Clean up oil spills (bioremediation) • Agriculture • Bt Corn ( pest resistant) • Food products • Cheese, Yogurt, Fermented foods, Bread, Alcohol) • Industry/manufacturing • Better cleaners
Insulin • The first commercial product made by genetic engineering (1976). • Insulin is the hormone used to control their blood sugar levels in diabetics • First sold in 1982, (S. San Francisco) PIGS E. Coli
Biotech Tools Restriction Enzyme: an enzyme that cuts DNA Ligase: an enzyme that pastes DNA back together • re-establishes the ester bond Gene of Interest: Example Insulin Gene Recipient Genome: E. coli plasmid • Plasmid: Circular DNA that bacteria have in addition to the normal chromosome
Restriction Enzyme Restriction Enzyme: an enzyme that cuts DNA • Natural function of restriction enzymes • To protect bacteria against viruses. • Digest foreign (e.g. viral) DNA. • Restriction enzymes are isolated from bacteria
A closer look…. EroR1 EcoR1 5’….ACTGTACGAATTCGCTA….3’ 3’….TGACATGCTTAAGCGAT….5’
A closer look…. EcoR1 AATTCGCTA….3’ GCGAT….5’ 5’….ACTGTACG 3’….TGACATGCTTAA “sticky ends” -can bind with other DNA molecules with the same overhangs
DNA Ligase DNA ligase 5’….ACTGTACAGATCCGCTA….3’ 3’….TGACATGTCTAGGCGAT….5’ DNA ligase
Let’s Try it • Genetic Engineering DEMO TAATGGATCCTT: Gene of Interest G GATCC BamH1 Sequence G AATTC EcoR1 Sequence • Crime Scene Activity • Tape: Ligase • Scissors: Restriction Enzyme
Create Your Own Recombinant DNA ! • Using the shorter sequence, create your plasmid, remember they are circular • Highlight the gene of interest TAATGGATCCTT • Figure our what restriction enzyme to use.GAATTC EcoR1 Sequence CTTAAG GGATCC BamH1 Sequence CCTAGG • The plasmid and DNA must be cut with same Restriction Enzyme • Create your Recombinant DNA.
HOW IS IT DONE?Steps for genetically engineering Insulin • Take a human cell and remove the nucleus • Extract the DNA (genome) from the nucleus • Locate the Insulin gene • Cut out the Insulin gene with a restriction enzyme • Take a Bacterial Plasmid and cut open with the same restriction enzyme • Place the Insulin gene into the bacterial Plasmid; you now have recombinant DNA. • Place the recombinant DNA back into an Bacterial Cell (transformation) • The Bacteria will now produce insulin.
Gel electrophoresis • We can use the fragments made by the restriction digestion to make a DNA fingerprint • DNA is negatively charged (due to phosphate backbone) • When we pump electrical charge into the buffer, it will make the DNA move towards the positive charge • Sorts DNA by length • Smaller DNA fragments travel further in the gel
Imagine you are a DNA molecule • If you were inside an agarose gel, your environment would resemble a very dense spider web. • The smaller the DNA fragment, the easier it is to get through the web. • This creates a DNA Fingerprint
Crime Scene DNA Who Committed The Crime?
Crime Scene Activity • Digest (Cut) the DNA samples with the restriction enzymes • Separate the fragments on the Electrophoresis Gel • Observe the differences and similarities to determine the guilty suspect.
Crime Scene Analysis • Restriction Digestion with EcoR1 (GAATTC) • Repeat with BamH1 (GGATCC) • Exclude Suspects • Plot fragments on Mock Gel (based on fragment sizes) • Answer questions. • Answer Wrap Questions • Find News article on DNA Fingerprinting