Genetic Engineering.pptx - MHSBioMrP2010

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Genetic Engineering

Chapter 10

What we are hoping to accomplish


Define genetic engineering.


Explain the principles of genetic engineering.


Explain and use the tools of genetic engineering.


Show and describe the practical uses of genetic
engineering.


Discuss the various steps in PCR and
transformation.

Biotechnology
-
Genetic Engineering


The application of biological and physical
principles to alter the genes of organisms to help
solve human problems


Improving crops and their yield


Producing insulin in bacteria


Genetically modified vaccines


Improving the freshness of fruits and vegetables


Biotechnology

Historical
Aspects

Plant breeding

Animal Breeding

Wine making

Dairy products

Bread

Antibiotics

Herbal medicine

Business and
Industry

Biomedical

Agriculture

Forensics

Industrial
production

Conservation

Wildlife
management

Multidisciplinary
Research

Biochemistry

Molecular genetics

Biophysics

Immunology

Cell culture

Fermentation

Bacteriology

Bioengineering

Genetic medicine

Conservation

Evolutionary
biology

Social and Ethical
Implications

Genetic privacy

Ownership of info.

Screening

Engineered food

Insurance

Treatment

DNA profiling

Patenting of life

What does genetic engineering mean?


It means to isolate a gene from the DNA of one
organism and transfer the gene into the DNA of
another.



We can do this because DNA is DNA in all
organisms. It is made out of the same 4 nitrogen
bases. The only difference is the sequence.



From there, DNA codes for RNA and proteins.

Recombinant DNA


A molecule of DNA made from pieces of DNA
from separate organisms.

Restriction Enzyme


An enzyme that is used to “cut” DNA at certain
places along
BOTH

sides of the DNA.



Most commonly used Restriction Enzymes


BamHI
-

G/GATTC


HindIII
-

A/AGCTT


HpaII
-

C/CGG


EcoR1
-

G/AATTC

Plasmids


Circular piece of DNA found in bacteria.


Replicates separate chromosome DNA


Contains very few genes


Useful because they have antibiotic resistance
genes on them.


Typically used as
VECTORS

to transfer DNA
from one organism to another.

Polymerase Chain Reaction (PCR)


A way to make an unlimited number of copies of
one gene.



How? DNA Polymerase is used to replicate the
same DNA segment over and over.



Why?


Increasing the number (Crime Scenes)


Sequencing (Prenatal diagnosis)


Cloning (Woolly Mammoth)

3 Steps to PCR

1.
Denaturing
-

Stands of DNA separate by
heating

2.
Annealing
-

Reaction cooled, primers bind

3.
Elongation
-

Reaction heated again, DNA
polymerase replicates the rest of the DNA from
primer.



Repeat steps 1
-
3 about 20
-
30x

DNA Fingerprinting


DNA is sliced into fragments called RFLPs.



DNA is then separated by sequence length by gel
electrophoresis.



The result is for an individual different lengths
of DNA that were cut by restriction enzymes that
varies among alleles and individuals.

Gel Electrophoresis

How the Gel Works?


DNA is negative



When DNA is placed next to a negative charge it
will want to move towards the positive charge.



The heavier strands will not travel as far as the
lighter strands.

DNA fingerprinting in a crime scene

Who did it? How Do We Know?

Crime Scene

Suspect 1

Suspect 2

Suspect 3

Suspect 4

Use in Crime Scenes


3 different Fingerprints are run



1/10,000 chance of matching an individual



Two fingerprints will only match 1 out of 10,000
X 10,000 times



Three matching prints would then be 1 in one
trillion


The DNA


Magnified using PCR



From blood, bone, flesh, hair follicle, semen,
saliva, any source of DNA



Analysis does not compare genes rather the
information between genes where a great
amount of variation exists


RFLP analysis


Restriction Fragment Length
Polymorphism (RFLP) Analysis


Identifies locations and numbers of restriction
sites of DNA.

Human Genome Project


Purpose: Sequence the entire human genome.


Why: detect, treat, and prevent genetic diseases.


Completed in 2003


Some of their findings for human DNA


Approximately 20,000
-
25,000 genes
(23,000)


3 billion base pairs make up the DNA.


Only 1.5% is actually coding for protein


Still left to do? Figure out how it all fits together.

Typical Transformation Experiment

1.
A target gene is isolated and removed by a
restriction enzyme
.

2.
Another restriction enzyme will cut the DNA of
a
vector

(
plasmids
) and insert our target
gene.

3.
Transfer the vector to the organism we want to
modify.

4.
Transformation
-

the gene becomes one with
its host and replicates producing clone copies.

5.
Screen the cells and select modified organism
by identifying our gene of interest.

Why do we care?

Genetically Engineered Drugs

Genetically Engineered Vaccines


Humulin

-

Diabetes


HGH
-

Dwarfism


Factor VIII, Factor IX
-

Hemophilia


t
-
PA


Heart attack, clot
buster


Lactoferrin



Milk protein



Herpes II


Hepatitis B


Influenza


Why do we care?

Cures for Genetic Diseases

Forensic Science


Cystic fibrosis


Genetic testing


Therapy


Karyotyping



DNA fingerprints


Crime


Paternity


Predisposed to disease


This may involve using stem
cells, by taking somatic cells
(like skin cells) and
dedifferentiating them down
to
pluripotent

stem cells.

Why do we care?

Plants

Animals


Ti plasmid causes plant
tumors


Glyphosate

resistant crops
(round
-
up ready)


Pest resistance


Improve taste and nutrition


Vaccines grown in plants



BST


Lactoferrin


Cloning