DNA TECHNOLOGY

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DNA TECHNOLOGY

• Transgenic organism
• Restriction Enzyme
• GMO genetically modified organism
• Gene therapy
• Vector
• Recombinant DNA
• PCR (polymerase chain reaction)
• Gel Electrophoresis
• Cloning
• Stem cells

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Transgenic Organisms
What are they?
Organisms that carry genes from another species
The first transgenic organisms were bacteria
First transgenic animal happened in 1975 A mouse
carried an ape gene

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How in the world did that happen?!
First you find the gene that you want to
transfer over
Then you must cut that gene out of the parent DNA
The area you want to put the DNA in the other
organism has to be found
Cut the new DNA so you can insert into vector

Gene put into its new DNA home
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Different restriction enzymes cut DNA at
different palindromes
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How do you cut the DNA?
Use restriction enzymes Bacterial proteins that
have the ability to cut strands of DNA at a
specific nucleotide sequence.
How does the cut piece of DNA get into the
targeted organisms cells?
For animals Transgene put into a vector and then
into a fertilized egg
For Plants Transgene put into vector which is
put into a bacterium that infects plant cells.
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What the heck is a vector?
A Vector is a Virus that has been re-engineered
A new piece of DNA has been added to the virus
and often times the part of DNA that infects you
is removed.
Transgene
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Transgene put into vector DNA
TTAAGTCAA
Egg put into mother and when baby cow is born it
makes human insulin
Vector puts Transgene into egg DNA
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Vectors

• Example plasmid (small ring of DNA in a
  bacterial cell
• May be biological (viruses or plasmids)
• Or mechanical (micropipette or bullet

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So why dont we make all kinds of new animals and
plants?
We dont know how it will affect our environment
If something is wrong with them its not like we
can just take them all back up
What other effects will it have besides the one
intended
They can mutate
Examples of GMOs (genetically modified
organisms) that we have now
Sterile male crop pests
Plants that have an insecticide in them
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Gene Therapy
What in the world is gene therapy?!?!
The treatment of certain disorders, especially
genetic disorders, by introducing specific
engineered genes into a patient's cells
Several methods can be used when treating a
genetic disorder
A normal gene may be inserted into a nonspecific
location within the genome to replace a
nonfunctional gene. This approach is most
common.An abnormal gene could be swapped for a
normal gene.The abnormal gene could be
repaired, which returns the gene to its normal
function.The regulation (the degree to which a
gene is turned on or off) of a particular gene
could be altered
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So how do we do this?
A vector is used to deliver the DNA needed to fix
the problem into the target cellsthe ones that
need to be fixed.
Just like in transgenic organisms the vector
infects the cells and delivers the DNA into the
cell to be put into the target cells DNA
This new DNA changes the target cells so that
they are now normal.
What kind of disorders are we talking about here?
Hemophilia
Sickle Cell Anemia
Cystic Fibrosis
Huntingtons Disease
Thalassaemia
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So dont we cure everybody?
FDA has not approved gene therapy because it has
proven dangerous.
Not a permanent cure
Our bodies can have an immune response to the
vector and DNA. Example of this is France.
Many diseases are caused by multi-gene problems
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Gene Therapy
Cell culture flask
Cell culture flask
Cell
Bone marrow cells
Hip bone
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PCR polymerase chain reaction

• Replicates DNA outside of living organisms
• Uses heat and enzymes to make lots of DNA very
  fast
• Used for crime investigations, diagnose diseases
  (like HIV) and cloning

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Recombinant DNA

• Made by connecting (recombining) fragments of DNA
  from different sources

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Uses of Recombinant DNA

• Insect resistant crops
• Growth hormones
• Insulin for diabetics
• Higher yielding food plants
• Clotting factor for hemophiliacs
• E. coli produces indigo dye to color jeans
• Cheese, laundry detergents, and sewage treatment
  are enhanced by recombinant DNA

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GEL ELECTROPHORESIS
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Loading the well with DNA
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Steps of Gel Electrophoresis

• Restriction enzymes cut DNA into fragments. The
  fragmented DNA is injected into wells in the gel.
  A current is sent through the gel and the
  fragments will move at different speeds that
  appear as bands under UV light. Bands can be
  matched up to identify criminals, bodies or
  fossils or to determine parentage. Longer
  fragments move slower than short ones.

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Send a current through the gel
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Fragments will move at different speeds, creating
bands that appear under UV light
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Bands can be matched up to identify parents,
children, or criminals
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Cloning

• Making an identical genetic copy

Dolly, 1997
News report
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Stem Cells

• Undifferentiated cells that can be genetically
  engineered to express the genes for any desired
  type of tissue
• Usually embryonic controversial
• But is there another way?
• Often used in regenerative medicine
• Current research hopes to treat diseases such as
  Parkinson's disease, Alzheimer's disease, spinal
  cord injury, heart disease, diabetes and
  arthritis.

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Plasmid Activity p. 355 in textbook

• Write the letters on the LONG strip
• GGATCC GGATCC
• CCTAGG CCTAGG
• Write the letters on the SHORT strip
• GGATCC
• CCTAGG
• Make sure all letters are equally spaced

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Plasmid activity p. 354-355

• Tape the ends of the shorter strip together.
• This is the plasmid
• Cut the longer strip of DNA in 2 places at the
  palindrome GIGATCC. This is the foreign DNA.
  Cutting it represents restriction enzymes.
• Cut the plasmid in the same way.
• Insert the foreign gene into the plasmid and tape
  together.

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