Genetic Engineering

Principles of
Genetic Engineering
 What is genetic engineering
• Genetic engineering, also known as
recombinant DNA technology, means altering
the genes in a living organism to produce a
Genetically Modified Organism (GMO) with a
new genotype.
• Various kinds of genetic modification are
possible: inserting a foreign gene from one
species into another, forming a transgenic
organism; altering an existing gene so that its
product is changed; or changing gene
expression so that it is translated more often
or not at all.
Basic steps in genetic engineering

1. Isolate the gene

2. Insert it in a host using a vector
3. Produce as many copies of the host as
possible
4. Separate and purify the product of
the gene
Step 1: Isolating the gene
Step 1: Alternative method (using
reverse transcriptase)

• Reverse transcriptase
• mRNA converted into cDNA
• Complementary strand produced using
DNA polymerase
• Advantage – more mRNA in cell than
DNA
 Step 2: Inserting gene into
vector
• Vector –
molecule of
DNA which is
used to carry a
foreign gene
into a host cell
Step 3: inserting vector into host
• See worksheet
Replica plating
Step 4: Multiplication of the host
cells by cloning

• Large scale fermenters by cloning

• All genetically identical because of
asexual reproduction
Step 5: Extraction of desired
gene product
• Genetic engineering: Changing the DNA in
living organisms to create something new.

• This organisms are called Genetically

Modified Organism (GMO)

• Example:
• Bacteria that produce human insulin

• Genetically Modified organism are called

transgenic organism; since genes are
transferred from one organism to another.
Some genetic engineering techniques are
as follows:
1. Artificial selection
A. selective breeding
B. hybridization
C. inbreeding

2. Cloning
3. Gene splicing
4. Gel electrophoresis: analyzing DNA
1. artificial selection: breeders choose
which organism to mate to produce
offspring with desired traits.

• They cannot control what genes are passed.

• When they get offspring with the desired
traits, the maintain them.

Three types of artificial selection:

A. selective breeding
B. hybridization
C. inbreeding
A. Selective breeding: when animals with
desired characteristics are mated to
produce offspring with those desired
traits.
• Passing of important genes to next
generation.
• Example: Champion race horses, cows
with tender meat, large juicy oranges on
a tree.
• For example people breed dogs for specific
purposes.
• Dachshund were once bred to hunt badgers
and other burrowing animals.
• They must be small to fit into the animals
hole in the ground.
• Selective breeding occurs when you choose
the best male and female to breed.
• This allows you to fine tune and control the
traits
• The offspring or babies will then have the
best traits.
• Then you continue to breed those organism
with the best traits, those traits will be
maintained.
• Examples of
selective breeding:

• Angus cows are bred to

increase muscle mass so
that we get more meat,

• Egg-Laying Hen-
produces more eggs
than the average hen
• B. Hybridizations: two individuals with unlike
characteristics are crossed to produce the best in
both organisms.
• Example: Luther Burbank created a disease
resistant potato called the Burbank potato.
• He crossed a disease resistant plant with one that
had a large food producing capacity.
• Result: disease resistant plant that makes a lot of
potatoes.
Other Examples of hybridization:
1. Liger: lion and tiger mix
2. Grape + apple= grapple. The fruit
tastes like grapes and looks like
apple.
C. Inbreeding breeding of organism that
genetically similar to maintain desired
traits.
• Dogs breeds are kept pure this way.
• Its how a Doberman remains a
Doberman.
• It keeps each breed unique from others.
• Risk: since both have the same genes,
the chance that a baby will get a
recessive genetic disorder is high.
• Risks: blindness, joint deformities.
• Variation: difference
between individuals of a
species.
• The differences are in the
genes but we see the
physical differences.
• For example: Some humans
have blond hair and some
have brown. This is a
variation among humans.
• Some finches have short
beaks, some have long
beaks.
• Inbreeding decreases
2. Cloning: creating an organism
that is an exact genetic copy of
another.
• There are human clones in our
school.
• identical twins are naturally
created clones.
• Clone: group of cells or
organisms that are genetically
identical as a result of asexual
reproduction
• They will have the same exact
How is cloning done?
► A single cell is removed from a parent
organism.
► An entire individual is grown from that
cell.
► Remember one cell has all the DNA
needed to make an entire organism.
► Each cell in the body has the same DNA,
but cells vary because different genes
are turned on in each cell.
 Dolly:
• Dolly was the first
mammal cloned.
• She had the same
exact DNA as her
mother and had no
father.
• Cloning is a form of
asexual reproduction.
• Only one genetic
parent.
http://content.tutorvista.com/biology_11/content/media/cloning.swf
• Since Dolly, cats and other organisms
have been cloned.
• The cat that was cloned had the same
exact DNA but different color fur than
the mother.

• How can this be?

• Environment plays a huge part in the way
organisms develop.
• Eggs are haploid
• Haploid: half the
chromosomes, 23 in
humans

• Body cells are diploid:

• Diploid: two sets of
chromosomes, one from
mom and one set from
dad 46 in humans.
How could you clone
a human?
• Step 1: An egg is
removed from a
female human
• Eggs are haploid: 23 23
chromosomes.
• The nucleus of the egg
EGG CELL

is removed and is
thrown away.
• Step 2: A body cell is
removed from
another person.
• The nucleus of the
body cell is removed
• Body cells are
diploid: 46
chromosomes. 46

Body Cell
• Step 3:
• The nucleus of the
diploid body cell is put
into the egg.
• This egg no longer
needs to be fertilized
since it has all 46 46
chromosomes.
EGG CELL
• Step 4: The egg is then
charged with electricity
to start mitosis.
• Step 5: Its then put into
a surrogate mother so it
can grow.

• Its going to be genetically

identical to the parent of
the body cell.
• But it will be a baby.

• Plants and animals can be

cloned.
Click and clone

• http://learn.genetics.utah.edu/content/tech
/cloning/clickandclone/
Benefits of cloning:
1. you can make exact
copies of organisms
with strong traits.
2. Increase food supply
3. Medical purposes: Saber Tooth Tiger extinct
clone organs for
transplants.
4. Bring back or Stop
species from going
extinct.
Risks of cloning:
1. Decreases genetic
diversity
2. If one of your clones
gets a disease, they all
get it: same immune
system.
3. Inefficient: high
failure rate: 90%+
4. Expensive
3. Gene splicing: DNA is cut
out of one organism and put
into another organism
• A trait will be transferred
from one organism to
another.
• For example: the human
insulin gene can be removed
from a human cell.
• It can be put into a
bacterial cell.
• The bacterial will now make
• This picture represents gene splicing.
• However, DNA is much smaller.
• Its done with high tech lab equipment
since DNA, is too small to hold or see
without a microscope.
The red piece the woman
is holding is an insulin
gene from a human
being. It is being
combined with DNA from
a bacteria.
Creates recombinant
DNA, something that has
never existed before.
Benefits:
• insulin is cheaper
• There are no side
effects because it
is human insulin.
• We once used pig
insulin but there
are side effects
and it more
expensive.
How are genes cut for gene
splicing?
• A bacterial plasmid is used.
• Plasmid: circular DNA in a bacteria
cell.
• It is very simple and easy to
manipulate.
• A restriction enzyme: enzyme that cuts the
DNA at a specific code.
• There are thousands of restriction enzymes.
• Each cuts DNA at a different sequence.
• Some look for GGCC and cut in between the
G and C.
• Every time GGCC is found in the DNA it is
cut by the restriction enzyme

DNA Code:

• TTATGGCCATACGGCCTT
• AATACCGGTATGCCGGAA
• TTATGGCCATACGGCCTT
• AATACCGGTATGCCGGAA

• TTATGG CCATACGG CCTT

• AATACC GGTATGCC GGAA

• This DNA segment was cut twice creating

three fragments.
• Since every one is different, we all have a
different amount of times GGCC is found.
• My DNA may be cut seven times
• Yours may be cut ten times.
This is how a restriction enzyme
works
How is gene
splicing done?
1. A restriction
enzyme cuts the
insulin gene out of
the human DNA.

2. A plasmid is
removed from a
bacteria and cut
with a restriction
enzyme
3. The human gene is place into the bacteria
plasmid
4. The plasmid is placed back into the bacteria.
• The cell now has directions (DNA) to make
insulin.
• That's exactly what it does.
• Its human insulin, bacteria do not make
insulin on their own.

Plasmid with
insulin gene
• This is called transformation: when a gene
from one organism is transferred to
different organism.
• The organisms that have DNA transferred
to them are called transgenic organisms.

• trans: means different,

• genic: refers to genes

• Genetic engineering has given rise to a new

technological field called biotechnology
(technology of life).
1. Transgenic (GMO) animals: genes
inserted into animals so they produce
what humans need.
• Why?: A way to improve the food
supply:
A. Transgenic cows: gene inserted to
increase milk production.
B. Spider goat: gene from spider
inserted into goat.
• Goats makes silk of the spider web in
their milk.
• Flexible, stronger than steel. Used in
bullet proof jackets.
C. Glow-in-the-
dark cats
• Scientist used a
virus to insert
DNA from
jellyfish
• The gene made
the cat produce a
fluorescent
protein in its fur.
2. Transgenic bacteria: gene inserted
into bacteria so they produce things
humans need.
• For example: insulin and clotting
factors in blood are now made by
bacteria.
3. Transgenic plants: plants are given
genes so they meet human needs.

A. Transgenic corn: given a gene so corn

produces a natural pesticide.
Now they don’t have to be sprayed with
cancer causing pesticides.
• 25% of all corn is like this.
B. Venomous cabbage
• gene from a scorpion tails
inserted into cabbage.
• Cabbage now produces that
chemical.
• Why? Limit pesticide use
while still preventing insects
from damaging crops.
• Corporations state the toxin
is modified so it isn’t
harmful to humans.
C. Banana vaccines
• virus is injected into a banana,
the virus DNA becomes part of
the plant.
• As the plant grows, it produces
the virus proteins — but not the
disease part of the virus.

• When people eat a bite, their

immune systems creates
antibodies to fight the disease
— just like a traditional vaccine

• Vaccines for hepatitis and

cholera
• A virus is often used to deliver DNA.
• In the movie “I Am Legend,” A healthy gene was
inserted into a virus.
• The virus invaded the cancer cells and inserts
the healthy gene to cure cancer.
• Worked at first but the virus mutated and
became deadly.
• This is being attempted in real life.
• Gene therapy: when disease causing
genes are cut out and good gene are
inserted.
• Restriction enzymes are used to cut out
bad genes.
• Viruses are used to insert good genes.
• Not approved for human use yet.
• Some possible side effects.
4. Gel electrophoresis: a
technique used to
compare DNA from two
or more organisms.

Why compare DNA:

1. Find your baby’s daddy
2. Who committed a crime.
3. How closely species are
related.
How is
electrophoresis
done?
A. The DNA is cut into
fragments with a
restriction enzyme.
B. The cut DNA is
then put into the
wells of a machine
filled with gel.
• The gel is spongy and
the DNA squeezes
through the pores.
C. The machine is plugged in and the fragments
get separated based on their size.
• The smaller fragments move further than the
large.
 Separation of DNA based on
size of fragments.

• Electrophoresis
results

Final result of electrophoresis

• Electricity provides the energy

• Why does DNA move?

• DNA has a negative charge.

• When the machine is plugged it, its moves towards

the positive pole created by the electricity
electrophoresis
Your DNA is so unique its considered to be
a DNA fingerprint.
Gel electrophoresis will separate your
DNA differently from anyone else.

Nova: who done it

http://www.pbs.org/wgbh/nova/sheppard/analyze.html

http://www.teachersdomain.org/asset/tdc
02_int_creatednafp2/
• Genetic engineering creates organisms
with recombinant DNA.
• Recombinant DNA: when DNA is
combined from at least two organisms.

Which techniques create recombinant DNA

1. Sexual reproduction: natural
2. selective breeding
3. Hybridization
4. Gene splicing
• Does cloning create organisms with
recombinant DNA?
• No, the DNA from one organism is
copied.
• DNA is not recombined.