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Ch.

13 Genetic Engineering

Ch. 13 Outline
13-1:

Changing the Living World

Selective

Breeding
Increasing Variation
13-2:

Manipulating DNA

The Tools

of Molecular Biology
Using the DNA Sequence

Ch. 13 Outline
13-3:

Cell Transformation

Transforming

Bacteria
Transforming Plant Cells
Transforming Animal Cells
13-4:

Applications of Genetic Engineering

Transgenic
Cloning

Organisms

What is genetic engineering?


In

1973, Mr. Cohen and Mr. Boyer


conducted an experiment on the DNA of
an American frog. They found and
isolated the gene that codes for ribosomal
RNA in the DNA of the frog. They
removed that gene from the frog and
inserted it into some E. Coli Bacteria.

What Happened?
During

transcription, the bacteria then


produced the frog RNA!

Genetic

Engineering: the process of


manipulating (moving) genes for practical
purposes (useful)
Recombinant DNA: DNA made from 2 or
more organisms that are different.

The Basic Steps of Genetic


Engineering
1.

Cutting the DNA:

Restriction Enzymes: bacterial enzymes that


recognize and bind to specific short
sequences of DNA, and then cute the DNA
between specific nucleotides within the
sequences.

Vector: agent used to carry the gene of


interest usually plasmids

Plasmid: the circular DNA molecules that


replicate

The Basic Steps to Genetic


Engineering
2.

Making Recombinant DNA

3.

DNA fragments of interest (that have already


been cut) are combined with the vector.
DNA ligase the enzyme bonds the 2 ends
of the fragments to the vectors.

Cloning

Gene cloning: the process of making many


copies of a gene

Bacteria reproduce by binary fission

The Basic Steps to Genetic


Engineering
4.

Screening

Cells that have received the gene of interest


are separated out.
Those cells then continue to produce the
protein coded for by the gene

Cutting DNA & Making


Recombinant DNA
How

Restriction enzymes work:

The

Enzymes recognize specific sequences on


Human and Bacterial Plasmids
The Enzymes cut the strands.
The cut produces DNA fragments with short strands
on each end that are complementary to each other
Sticky

Ends

Both

the human DNA and the Plasmid Open Up


with the same sticky ends remaining
They

Bind Together

Diagram
Recognition sequences

DNA sequence

Recognition sequences

DNA sequence

Restriction enzyme
EcoRIcuts the DNA
into fragments.

Sticky end

Confirmation of a Cloned Gene

One method used identify a specific


gene is called a Southern Blot
Steps:
1.
2.

Cut DNA from bacteria with restriction


enzymes.
DNA fragments are separated by a gel
soaked in a chemical solution.

Gel electrophoresis uses an electric field


within a gel to separate molecules by their size

Confirmation of a Cloned Gene


Negatively

charged DNA is put into these

wells.
They

are attracted to the positive pole from


the electric field.

The

Smallest DNA fragments move the


fastest

Gel Electrophoresis
Power
source

DNA plus restriction


enzyme

Longer
fragments
Shorter
fragments
Mixture of DNA
fragments

Gel

Confirmation of a Cloned Gene


3.

The DNA separated is then transferred to


a filter paper (blotted) and a probe
solution is added.

4.

Probes: radioactive RNA or single-stranded


DNA pieces that are complementary to the
gene of interest

Only DNA fragments complementary to


the probe will form and bind bands

Confirmation of Cloned Genes


Why

do this?

Bacterial

colonies can be used to produce


large quantities of the protein (used to study
or make drugs)

Genetically engineered Drugs and


Vaccines
Today, many

pharmaceutical companies
around the world produce important
proteins using genetic engineering.
Vaccine: a solution containing all or part
of a harmless version of a pathogen; used
to prevent viral diseases (dont respond to
drugs)
Many vaccines are made using genetic
engineering

DNA fingerprinting
DNA fingerprinting:

a pattern of dark bands on


photographic film that is made when an
individuals DNA restriction fragments are
separated by gel electrophoresis, probed, and
exposed to X-ray film.
DNA fingerprints can be used to establish
paternity, identify genetic disorders, or in
forensics (scientific study of cause of injury or
death in criminal activity)

Improving Crops
Genetic

engineers can add favorable


characteristics to a plant
Resistant

to insects (no longer need


pesticides); resistant to weed killer (so crops
wont die, but weeds will); improved nutrition
rice + corn

Plant Transformation
Agrobacterium
tumefaciens
Gene to be
transferred

Cellular
DNA

Recombinant
plasmid

Inside plant cell,


Agrobacterium inserts part of
its DNA into host cell
chromosome

Plant cell colonies

Transformed bacteria introduce


plasmids into plant cells

Complete plant is
generated from
transformed cell

Animal Farming
Growth

hormones given to cows to


produce more milk
Human genes added to farm animals in
order to have human proteins in their milk
The

Human proteins are extracted from milk


and sold to pharmacy companies.
Useful

for complex proteins that cant be made in


bacteria

Creating HGH
Recombinant
DNA

Gene for human


growth hormone

Gene for human


growth hormone
Human Cell

Sticky
ends

Bacterial Cell

DNA
recombination
DNA
insertion

Bacterial
chromosome

Plasmid

Bacterial cell for


containing gene for
human growth hormone

Animal Farming
Transgenic

animals: Animals that have foreign


DNA in their cells
Cloning of animals is another way to make large
quantities of a certain protein.
How it works: an intact nucleus from an
embryonic cell (whose DNA has recombined
with a human gene) is placed into an egg whose
nucleus has been removed. The new egg is
then placed into the uterus of an animal.

Cloning
A donor cell is taken from
a sheeps udder.

Donor
Nucleus
These two cells are fused
using an electric shock.

Fused Cell

Egg Cell
The nucleus of the
egg cell is removed.
An egg cell is taken
from an adult
female sheep.

The fused cell


begins dividing
normally.

Embryo

Cloned Lamb
The embryo
develops normally
into a lambDolly

Foster
Mother

The embryo is placed


in the uterus of a foster
mother.