What Is Biotechnology? Any technique that uses living organisms or substances from those organisms...
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Transcript of What Is Biotechnology? Any technique that uses living organisms or substances from those organisms...
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
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?
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
GMO’s: Transgenic Organisms: Bt Corn ( pest resistant) Recombinant DNA (rDNA): rInsulin
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
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
Biotech Tools
Restriction EnzymeRestriction Enzyme: an enzyme that cuts DNALigaseLigase: an enzyme that pastes DNA back together
re-establishes the ester bondGene of InterestGene of Interest: Example Insulin GeneRecipient GenomeRecipient Genome: E. coli plasmid
Plasmid: Circular DNA that bacteria have in addition to the normal chromosome
Restriction EnzymeRestriction Enzyme
Natural function of restriction enzymes To protect bacteria against viruses. Digest foreign (e.g. viral) DNA. Restriction enzymes are isolated from
bacteria
Restriction EnzymeRestriction Enzyme: an enzyme that cuts DNA
A closer look…. EroR1
5’….ACTGTACGAATTCGCTA….3’3’….TGACATGCTTAAGCGAT….5’
EcoR1
A closer look…. EcoR1
AATTCGCTA….3’
GCGAT….5’5’….ACTGTACG
3’….TGACATGCTTAA
“sticky ends”-can bind with other DNA molecules with the same overhangs
5’….ACTGTACAGATCCGCTA….3’ 3’….TGACATGTCTAGGCGAT….5’
DNA Ligase
DNA ligase
DNA ligase
Create Your Own Recombinant DNA !
1. Using the shorter sequence, create your plasmid, remember they are circular
2. Tape= DNA Ligase ; Scissors= restriction enzymes
3. Highlight the gene of interest TAATGGATCCTT4. Figure our what restriction enzyme to use.
GAATTC EcoR1 Sequence CTTAAGGGATCC BamH1 SequenceCCTAGG
4. The plasmid and DNA must be cut with same Restriction Enzyme5. Create your Recombinant DNA.
HOW IS IT DONE?Steps for genetically engineering Insulin
1. Take a human cell and remove the nucleus2. Extract the DNA (genome) from the nucleus3. Locate the Insulin gene4. Cut out the Insulin gene with a restriction enzyme5. Take a bacterial plasmid and cut open with the same
restriction enzyme6. Place the Insulin gene into the bacterial plasmid; you
now have recombinant DNA.7. Place the recombinant DNA back into an bacterial cell
(transformation)8. The bacteria will now produce insulin.
A closer look…. HindIII
5’….ACTGTACAAGCTTGCTA….3’3’….TGACATGTTCGAACGAT….5’
HindIII
A closer look…. HindIII
AGCTTGCTA….3’
ACGAT….5’5’….ACTGTACA
3’….TGACATGTTCGA
“sticky ends”-can bind with other DNA molecules with the same overhangs
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.
Famous DNA Cases
The Romanov's and Anastasia OJ Simpson (DNA proved guilt but people
didn’t 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 wasn’t available at the time
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
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/
content/gelelectrophoresis.html
Crime Scene DNA
Who Committed The Crime?
Famous DNA Case
Let’s Try it
Crime Scene DNAGAATTCTAATGGATCCTT
G AATTC EcoR1 SequenceG GATCC BamH1 Sequence
Creates 3 pieces1 bp , 10 bp, 7 bp
10 bp
7 bp
1 bp
+
_
Lab: Crime Scene Analysis
1. Restriction digestion with BamH1 (GGATCC) Use blue pen
2. 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
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?v=bbZiOiPVG6c
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.
Let’s Click and Clone
http://learn.genetics.utah.edu/content/tech/cloning/clickandclone/