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/