What Is Biotechnology?

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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

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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?

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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
• GMOs
• Transgenic Organisms Bt Corn ( pest resistant)
• Recombinant DNA (rDNA) rInsulin

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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

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rInsulin

• 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
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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

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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

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A closer look. EroR1
EcoR1
5.ACTGTACGAATTCGCTA.3 3.TGACATGCTTAAGCGAT.5

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A closer look. EcoR1
AATTCGCTA.3 GCGAT.5
5.ACTGTACG 3.TGACATGCTTAA
sticky ends -can bind with other DNA molecules
with the same overhangs
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DNA Ligase
DNA ligase
5.ACTGTACAGATCCGCTA.3
3.TGACATGTCTAGGCGAT.5
DNA ligase
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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.

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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.

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A closer look. HindIII
HindIII
5.ACTGTACAAGCTTGCTA.3 3.TGACATGTTCGAACGAT.5

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A closer look. HindIII
AGCTTGCTA.3 ACGAT.5
5.ACTGTACA 3.TGACATGTTCGA
sticky ends -can bind with other DNA molecules
with the same overhangs
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Restriction Enzymes DNA Evidence

• Original Cases Timeline

• 1984 Alec Jeffreys develops DNA fingerprinting,
  for forensics and paternity cases.
• 1987 Tommie Lee Andrews is convicted of rape in
  Orlando based on DNA fingerprinting--the first
  such case in the nation.
• 1989 DNA evidence for the first time overturns a
  conviction, clearing Gary Dotson of an Illinois
  rape.

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Famous DNA Cases

• The Romanov's and Anastasia
• OJ Simpson (DNA proved guilt but people didnt
  understand DNA evidence)
• Thomas Jefferson and Sallie Heming's kids
• Use to identify 9-11 remains
• Innocence Project to date 305 wrongful
  convictions overturned by DNA evidence, that
  wasnt available at the time

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Gel electrophoresis

• Cuts DNA at repetitive DNA that everyone has but
  are unique called Restriction Fragment Length
  Polymorphisms (RFLP)
• We can use the fragments made by the restriction
  enzyme digestion to make a DNA fingerprint
• Looking for 13 variations
• 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

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Imagine you are a DNA molecule

• Sorts DNA by length
• Smaller DNA fragments travel further in the gel
• If you were inside an agarose gel, your
  environment would resemble a very dense web.
• The smaller the DNA fragment, the easier it is to
  get through the web.
• This creates a unique DNA banding pattern called
  a DNA Fingerprint

http//www.sumanasinc.com/webcontent/animations/co
ntent/gelelectrophoresis.html
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Crime Scene DNA
Who Committed The Crime?
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Famous DNA Case
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Lets Try it

• Crime Scene DNA
• GAATTCTAATGGATCCTT
• G AATTC EcoR1 Sequence
• G GATCC BamH1 Sequence
• Creates 3 pieces
• 1 bp , 10 bp, 7 bp

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Lab Crime Scene Analysis

• Tape DNA Ligase Scissors restriction enzymes
• Restriction digestion with BamH1 (GGATCC)
• Use blue pen
• Restriction digestion with EcoR1 (GAATTC)
• Use red pen
• Plot fragments on mock gel (based on fragment
  sizes)
• Exclude suspects
• Answer questions on mock gel WS
• Answer wrap questions
• Find news article on DNA fingerprinting and
  summarize article

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Somatic Cell Nuclear Transfer (SCNT)

• aka Cloning
• 1952 Xenopus Tadpoles
• 1996 The first mammal cloned from adult cells
  was Dolly, the sheep.
• 2001 The first human embryo cloned, but on
  divided to six cells, providing evidence that
  human cloning is not currently possible
• http//www.youtube.com/watch?vbbZiOiPVG6c

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Dolly 277 tries 1 Sheep
On 14 February 2003, Dolly was euthanized because
she had a progressive lung disease and severe
arthritis. Normal Life expectancy 11-12 yrs.
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Lets Click and Clone

• http//learn.genetics.utah.edu/content/tech/clonin
  g/clickandclone/