Genetic Engineering

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Genetic Engineering Biotechnology
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Major Techniques

• PCR
• Polymerase Chain Reaction
• Restriction Endonucleases and Gel Electrophoresis
• DNA Profiling/DNA Fingerprinting
• Gene Transfer Transformation

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Outline the use of Polymerase Chain Reaction
(PCR) to copy and amplify minute quantities of
DNA Specifics not required

• Used to make millions of copies of select section
  of DNA
• When small amount of DNA are found but large
  amounts are needed for analysis
• Semen, blood, other tissues, long-dead specimens
  DNA from all can be amplified
• Thermus aquaticus hot springs bacterium
• http//www.dnalc.org/ddnalc.org/resources/pcr.html
  

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STATE that in gel electrophoresis fragments of
DNA move in an electric field and are separated
according to their size
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• Separate DNA, proteins or other molecules based
  on charge and size

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STATE that gel electrophoresis is used in DNA
Profiling/Fingerprinting
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Describe the application of DNA Profiling to
determine paternity and also in forensic
investigations

• CSI
• Paternity
• CDC epidemiology disease transmission
• Geneology
• Breeding authenticity

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Analyze DNA profiles to draw conclusions about
paternity or forensic investigations
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STATE that,

• when genes are transferred between species, the
  amino acid sequence of the polypeptide translated
  from them is UNCHANGED because the genetic code
  is UNIVERSAL

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Outline three outcomes of the sequencing of the
complete human genome

• HOMEWORK!

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Organisms that have had genes transferred into
them from a different species are GMOs or
Transgenic Orgs.
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Outline a basic technique used for gene transfer
involving plasmids, a host cell (bacterium, yeast
or other cell), restriction enzymes
(endonucleases) and DNA ligase
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Outline a basic technique used for gene transfer
involving plasmids, a host cell (bacterium, yeast
or other cell), restriction enzymes
(endonucleases) and DNA ligase

• Messenger RNA coded for insulin is extracted from
  human pancreas
• DNA copies of the messenger RNA coding for
  insulin are made using the enzyme reverse
  transcriptase.
• Plasmids are small loops of DNA found in
  bacteria.
• Plasmids are cut open using restriction enzyme
  endonucleouse.
• The insulin gene and the plasmid are mixed.
• DNA ligase seals up the plasmid.
• The plasmid with the human insulin gene is called
  a recombinant plasmid.
• The recombinant plasmid are mixed with a strain
  of E.Coli bacteria.
• The E.coli bacteria that pick up the plasmid
  start to make insulin which is then extracted,
  purified and used by patients.

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State 2 examples of the current uses of GM-crops
or GM-animals

• Ex. Golden rice - produces beta-carotene, which
  can be metabolized into Vitamin A within the
  body. Scientists hope that it will eventually be
  a cheap source of beta-carotene in malnourished
  countries, reducing the number of children
  worldwide that go blind from Vitamin A
  deficiency.
• Bt maize is a genetically modified corn crop that
  produces a toxin that kills European corn borers
  feeding on the maize

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GM-animal examples
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Cats, Goats, Chickens
GM hens - produce cancer-fighting medicines in
their eggs. Human genes have been added to their
DNA so that human proteins are secreted into the
whites of their eggs, along with complex
medicinal proteins similar to drugs used to treat
skin cancer and other diseases.   What exactly do
these disease-fighting eggs contain? The hens lay
eggs that have miR24, a molecule with potential
for treating malignant melanoma and arthritis,
and human interferon b-1a, an antiviral drug that
resembles modern treatments for multiple
sclerosis.
Strong, flexible spider silk is one of the most
valuable materials in nature, and it could be
used to make an array of products from
artificial ligaments to parachute cords
.  Researchers inserted a spiders dragline silk
gene into the goats DNA in such a way that the
goats would make the silk protein only in their
milk. This silk milk could then be used to
manufacture a web-like material called Biosteel.
Researchers took skin cells from Turkish Angora
female cats and used a virus to insert genetic
instructions for making red fluorescent protein.
  Whats the point of creating a pet that doubles
as a nightlight? Scientists say the ability to
engineer animals with fluorescent proteins will
enable them to artificially create animals with
human genetic diseases
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Discuss the potential benefits and possible
harmful effects of one example of genetic
modificationHOMEWORK
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Define Clone

• Clone a group of genetically identical organisms
  or group of organisms derived from a single
  parent (in nature it is called Asexual
  Reproduction)

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Outline a technique for cloning using
differentiated animal cells

• Udder cells are taken from a
• donor sheep. The genes in the
• cell are made dormant.
• Unfertilized eggs are taken
• from another sheep.
• The nucleus is removed from
• the egg cell.
• The egg cell without a nucleus
• is fused with the udder cell
• using a pulse of electricity.
• The fused cells develop into
• zygotes.
• Inserted back into mother.

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Discuss the ethical issues of Therapeutic Cloning
(TC) in humans

• TC is the creation of an embryo to supply
  embryonic stem cells for medical use. The embryo
  does not survive the processonly its stem cells
  do.
• You should be able to discuss the ethical
  implications