Download DNA Technology

DNA
Technology
1
DNA
Technology
is used for
1. produce pharaceutical
products
2. produce herbicide
resistant crops
3. treat genetic diseases
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What is MOLECULAR
BIOLOGY?
• Bio on the molecular level
• Like GENETIC ENGINEERING
• -when we alter/manipulate genes
• For what reasons?
– Cure diseases/mutations (GENE
THERAPY)
– Improve food crops
– What else?!
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1. DNA Extraction
• Chemical treatments (like
adding detergents) cause cells
and nuclei to burst and
remove DNA from cells
• The DNA is inherently sticky,
and can be pulled out of the
mixture
• This is called “spooling” DNA
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“Spooled” DNA
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2. Splicing (cutting) DNA
• Restriction enzymes (ex.
nuclease) cut DNA at specific
sequences
• Divides DNA into manageable
fragments
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Sticky ends
-places on DNA where R.E. cut
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3. Transferring Genes
• Uses a PLASMID – is an extra
ring of DNA
– Ring of DNA in a bacterium (it’s
not the main chromosome tho)
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RECOMBINANT
PLASMID
-changed DNA
the plasmid acts
like a host for
the
cloning vector to
be placed inside
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How does the
prokaryote know
how to express
eukaryote
genes?
• AH! YOU MAKE
SURE YOU CUT
THE TARGET GENE
WITH ITS
PROMOTER!
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Making
insulin
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TRANSGENIC ORGANISM
-host organism that gets rDNA
(recombinant DNA)
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4. Gel Electrophoresis
• DNA fragments can
be separated based
on size
• The phosphate
groups on outside
backbone are
negatively charged
• DNA is placed in a
gel and electricity is
run through
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Gel Electrophoresis
• Negative DNA moves toward
the positive end
• Smaller fragments move
farther and faster
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What cuts DNA into smaller
chunks so they can drop out
throughout the gel?
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DNA FINGERPRINTING
by Gel Electrophoresis
• Pattern of
bands
made up
of specific
DNA
fragments
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Which suspect was at the
crime scene?!
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5. DNA
SEQUENCING
• Figure out the ORDER
of NUCLEOTIDES
(bases)
• Inject DNA with dyes
that will bind to specific
bases
• Bases will drop out at
certain known
“heaviness” spots so
you can sequence it!
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6. PCR - Copying DNA
• Polymerase Chain Reaction
• Also called PCR
• A method of making many quick
copies of a piece of DNA
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Steps in Copying DNA
• A DNA molecule is placed in
a small test tube
• +DNA polymerase
• +heat to separate into ss
• +primers (ss matching DNA
pieces to start replication)
• +nucleotides
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Copying DNA
• The tube is cooled, and
DNA polymerase adds new
bases to the separated
strands
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PCR
Large amounts of DNA can be made
from a small starting sample
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Dr. Kary Mullis
• 1993
• Dr. Kary Mullis
(1944) discovered
the PCR procedure,
while HIGH on LSD!
7. Cloning
• Clone- can produce
many genetically
identical cells or
offspring
• produced by
asexual
reproduction
(mitosis)
• We can quickly
produce organisms
with desirable
traits
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Cloning organisms
• A body cell from one
organism
• and an egg cell from
another
• are fused
• The resulting cell divides
like a normal embryo
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What was the first cloned organism?
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Dolly (born 1996) and her
surrogate Mommy
Who was
st
the 1
cloned
MAMMAL
?
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Dolly and
her baby,
Bonnie
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Normal sheep lifespan 10-12 yrs
Dolly died
in 2003
(put to
sleep after
seeing
lung
tumors)
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We clone animals…(lab
and domesticated)
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2001 We’ve brought back
an extinct animal – the
bucardo goat (but it died
of lung defect)
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List of animal clones
•
•
•
•
•
•
•
•
•
•
•
•
•
1 Carp •
2 Cat •
3 Cattle •
4 Deer •
5 Dog •
6 Ferret •
7 Frog (tadpole)
•
8 Fruit flies
•
9 Gaur •
10 Goat •
11 Horse•
12 Mice •
13 Mouflon
14 Mule
15 Pig
16 Pyrenean ibex
17 Rabbit
18 Rat
19 Rhesus Monkey
20 Sheep
21 Water Buffalo
22 Wolf
23 Zebrafish
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st
1
2007
primate stem
cells cloned
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2013
st
1
HUMAN stem
cells cloned
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CLONING =
DE-EXTINCTING ANIMALS
• MAMMOTH
• -overhunting
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GASTRIC-BROODING FROG
-habitat loss, pollution, disease?
(Australia)
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PASSENGER PIGEON
-overhunting (fed to slaves!)
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DODO
-overhunted by sailors
(Mauritius)
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8. Human Genome
Project
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Human Genome Project
• 1990 -2003
• Research effort to sequence
all of our DNA (46
chromosomes)
• Over 3 billion nucleotides
• Mapping every gene location
(loci)
• Conducted by scientists
around the world
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Benefits of Human
Genome Project
• Improvements in medical
prevention of disease, gene
therapies, diagnosis techniques …
• Production of useful protein
products for use in medicine,
agriculture, bioremediation and
pharmaceutical industries.
• Improved bioinformatics – using
computers to help in DNA
sequencing …
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9. Biotechnology •
TO create new
products from
ALTERING GENES in
plants and animals for
more desirable traits
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GM FOODS
-Genetically Modified
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Herbicide Resistant
Crops
+ CP4 EPSPS = Roundup
gene
Ready
•
•
•
•
Soybeans:
Corn:
Cotton:
Canola:
Roundup Ready
Roundup Ready, Liberty Link
BXN, Roundup Ready
Liberty Link, Roundup Ready
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Biotechnology Breakthroughs
• Rice
– Enriched with beta-carotene
and iron
• Bananas
– Containing edible hepatitis
vaccine
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Biotechnology Breakthroughs
• Potatoes with higher solid
content
• Garlic that lowers cholesterol
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GM vs Artificial Selection
Pros
• Reduced pesticide use
• Lower energy requirements
• Cleaner water
• Less soil erosion
• More food production!
• Cheaper! $
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Cons
• Not natural
• Does it cause health issues?
• Should it be labeled?
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•
•
•
•
Corn – insecticide
Soy – herbicide
Cotton – pesticide
Papaya – virusresistant
• Rice – Vitamin A
• Dairy – bovine GH
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10. Epigenetics – altering genes by
messing with OUTSIDE DNA stuff like
RNA or HISTONES!
Which mouse did we
methylate (add CH3
group to its histone
proteins that DNA
wraps around?
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11. iPSC
But…
induced Pluripotent Stem Cells
-turning BODY cells BACK into
STEM cells! (no embryo killing)
Transfect
with viral
vectors
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Causes cancer
bc virus doesn’t
know when to stop
telling cells to
reproduce
the rate at which
body cells are
reprogrammed into
stem cells is LOW!
0.1%! Bc we need
to know the
EXACT time,
balance and levels
of expression of
gene we want.
Stem cells are
undifferentiated cells
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