Biotechnology

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Biotechnology

• You Will Learn About
• Transformation
• Cloning
• DNA Fingerprinting by Restriction Fragment Length
  Polymorphism (RFLP)
• What is the name of the lamb?

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Brainstorm

• What are some ways in which DNA can be used as a
  tool (think about TV, the news)

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Goals of Applied Genetics

1. Help humans create crops that can be frost
   resistant (we love our peaches!)
2. Use transgenic organisms to help medical
   researchers model human physiology for testing
3. Help industry to create bacteria to break down
   pollutants into harmless products
4. Pharmaceutical companies use recombinant DNA to
   cheaply produce human hormones (insulin) and
   other proteins
5. Help solve crimes and determine relatives

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Technologies Addressed in this Showplease write
these on first slide

• Bacterial Transformation
• Cloning
• DNA fingerprinting (RFLP analysis)

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Selective breeding

• Selecting organisms with the most desirable
  traits
• Requires time and several generations to produce
  offspring with the desired trait
• Ex
• Short vs long haired cats
• Milk production in cattle
• Disease resistant foods
• Bacteria that break down oil

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Bacterial Transformation

• Also called recombinant DNA technology or gene
  cloning
• Uses a bacterial host because of fast
  reproduction and a circular DNA vehicle to hold
  the foreign DNAplasmid
• Organisms containing recombinant DNA or foreign
  DNA are known as transgenic

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Steps involved

1. Select the desired gene(s) to be inserted into
   the organism and a bacterial host containing a
   plasmid (vehicle to hold the desired gene)
2. Cut two DNA molecules into fragments with special
   (restriction) enzymes
3. Splice (rejoin) the fragments (gene and plasmid)
   together in the desired combination
4. Introduce or insert the new DNA into a living
   cell for replication (mitosis)

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Vector to transfer DNA
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Enzymes

• Restriction Enzymes Used to cut a DNA molecule at
  a specific nucleotide sequence
• Produces one of two types of DNA fragments
• Sticky ends (palindrome)
• Blunt ends

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Sticky Ends (palindrome)
Blunt ends
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Gene Cloning

• Used to produce genetically identical copies of a
  cell, tissue, organ, and/or organism
• Needed to produce multiple copies of the desired
  DNA

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Cloning Applications

• Currently
• Plants are cloned to produce a large number of
  genetically identical plants in a short amount of
  time
• Future
• Clone productive and healthy animals to increase
  yield for farmers and to grow organs for
  transplants

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How cloning works.
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Gene Therapy

• The insertion of normal genes into human cells to
  correct genetic disorders like cystic fibrosis.
  (Not successful yet?)

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DNA Fingerprinting(RFLP Analysis)

1. Obtain a small sample of DNA
2. Make millions of copies using PCR (polymerase
   chain reaction) technique
3. Cleave (cut) DNA with restriction enzymes
4. Separate DNA fragments using gel electrophoresis
   and compare
5. Each humans DNA will have some unique pieces
   because each of our DNA is unique

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Gel Electrophoresis

• Gel electrophoresis is used to create a DNA
  fingerprint
• Small amounts of DNA are loaded into wells in the
  gel.
• An electric current pushes the small pieces of
  DNA farther down the gel than the larger pieces.
• DNA patterns are compared to known patterns
  (Human Genome Project).
• Used for forensics, blood samples, and paternity
  tests (children get half their bands from mom and
  half from dad)

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Running a gel
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How DNA moves
Step 1 Make wells in gel.
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Step 2 Pipette DNA into wells.
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Step 3 Run an electric current to watch DNA
migrate.
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Step 4 Notice similarities and differences in
banding patterns.
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The Human Genome Project

• There are approximately 25,000 genes on the 46
  human chromosomes (far fewer than expected)
• Human Genome Project- an international effort to
  completely map and sequence human chromosomes
  (completed April 2003)
• This has been important in
• identifying genes that are related to disease.
• Determining human relationships with other
  species.

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Microarray

• An new technology, the microarray, allows a cell
  to be samples for many genes at once.
• It is involved in finding
• What genes are important in cancer and cancer
  prevention.
• What medicines can be used for a person
  (customized medicine)
• Much more, and we will spend some time with this.