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Genetic Engineering
- is a process of making changes on the genetic code of an organism. Its goal
is to add one or more new traits that are not normally found in that
organism. Through advanced studies in the structure of DNA and its chemical
properties, scientists have been able to employ different techniques to
extract, cut, and make unlimited copies of DNA.

The Importance And The Essence Of

Genetic Engineering
DNA recombination - is a process of
modifying the genes of organisms
for practical purposes. It is done
when a piece of DNA is combined
with another DNA from another
source. The resulting genetic
product is called recombinant DNA.
With this process, organisms get to
have traits that are not normally
found in their species.

Genetic Engineering Techniques

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, they maintain them.

Three types of Artificial Selection:

A. Selective breeding: when animals with desired characteristics are

mated to produce offspring with those desired traits.
● Passing important genes to the next generation.
● 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 organisms with the best
traits, those traits will be maintained.

B. Hybridizations: two individuals with unlike characteristics are

crossed to produce the best in both organisms.

C. Inbreeding: breeding of organisms that are genetically similar to

maintain desired traits.
 ● Risk: since both have the same genes, the chance that a baby
will get a recessive genetic disorder is high. (ex: 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 variations.

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 DNA as the parent.

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

How could you clone a human?

Step 1: An egg is removed from a female human

● Eggs are haploid: 23 chromosomes.

● The nucleus of the egg 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.

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

chromosomes.

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.
● It's going to be genetically identical to the parent of the body
cell but it will be a baby.
● Plants and animals can be cloned.
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 bacteria will now make human insulin.

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’s
more expensive.

Processes Used in Recombinant DNA Technology

There are three methods by which recombinant DNA is made. These are
transformation, vectorless gene transfer, and transduction.

1. Transformation Using a Vector: Recombinant DNA may be created

through transformation with the help of a vector, such as bacterial
cells. Vectors are organisms that are normally harmless but may help
spread infection by transferring the genetic material from one host to
another.
● In the transformation process, a selected portion of the foreign
DNA is inserted into a small, circular DNA molecule called plasmid,
which is naturally found in bacteria. Plasmids are the most useful
tool in gene transformation for two reasons. First, a plasmid
contains a gene sequence that serves as a bacterial origin of
replication. This is where the foreign DNA can be inserted into the
bacterial cell. Second, it also contains a genetic marker, which
makes it possible to distinguish bacteria that carry the
plasmid-containing foreign DNA. Some of these genetic markers
code for antibiotic resistance.

How are genes cut for gene splicing?

● A bacterial plasmid is used.

1.1 Plasmid: circular DNA in a bacteria cell.

● It is very simple and easy to manipulate.

1.2 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
 ● This DNA segment was cut twice creating three fragments.
● Since everyone is different, we all have a different amount of
times when GGCC is found.
● My DNA may be cut seven times
● Yours may be cut ten times.

During transformation, a restriction endonuclease enzyme is used to cut

the piece of the donor DNA. This enzyme cleaves the DNA at the
phosphate-sugar bond, and thus sticky ends are created. Sticky ends
are areas in the DNA where the bases are ready to be paired.
Restriction enzymes cut the DNA only at a specific nucleotide sequence.
They work precisely like a key that fits only one specific lock. Then, an
enzyme known as DNA ligase is used to insert the donor DNA into the
vector. It seals the sticky ends by joining the phosphate and the sugar
bonds in the
DNA. The inserted DNA also contains a genetic marker for identification.
The recombinant DNA is then inserted into a bacterial cell, such as E.
coli.

● The organisms that have DNA transferred to them are called

transgenic organisms.

Transgenic (GMO) animals: genes inserted into animals so they produce

what humans need.
Ex:
Transgenic cows: genes inserted to increase milk production.
Spider goat: gene from spider inserted into goat.

● Goats can make silk of the spider web in their milk.

● Flexible, stronger than steel. Used in bullet proof jackets.

Transgenic bacteria: genes inserted into bacteria so they produce things

humans need.

Ex: insulin and clotting factors in blood are now made by bacteria.

Transgenic plants: plants are given genes so they meet human needs.

Ex:

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.

Gene therapy: when disease causing genes are cut out and good
genes are inserted.

● Restriction enzymes are used to cut out bad genes.

● Viruses are used to insert good genes.
● Not approved for human use yet.
● Has some possible side effects.
2. Vectorless Gene Transfer: This process is similar to transformation, but
it does not involve vectors. The types of vector less gene transfer
include electroporation, protoplast fusion, microinjection, and use of a
particle gun.

Types of Vectorless Gene Transfer

● Electroporation: temporary holes are formed in the plasma

membrane of the host cell by applying a significant amount of
electricity in the culture medium. This enables the entry of foreign
DNA via the pores.

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.

● Protoplast Fusion: Cells are treated with chemicals to initiate

recombination. In this process, bacterial cell walls are digested,
turning the cells into protoplasts. These protoplasts are treated
with polyethylene glycol to allow them to fuse, creating a random
recombination of genes. The resulting recombinant cell will now
grow a new cell wall.

● Microinjection: The host cell is immobilized by applying a mild suction

with a blunt pipette. The foreign gene is then injected with a
microinjection needle, thus creating recombinant DNA.

● Particle Gun: the host cell is bombarded with tungsten particles

coated with foreign DNA. This process is used in the field of
agriculture. Many farmers use this method to genetically modify
plants to make them highly resistant to insects and other pests.
Some also use this method to develop crops that can survive
extreme weather conditions. Electricity provides the energy

Why does DNA move?

● DNA has a negative charge.
 ● When the machine is plugged in, it moves towards the
positive pole created by the electricity.

3. Transduction: the process wherein genetically engineered bacterio

phages viruses that parasitize bacteria- are introduced into the cell to
create the desired recombinant DNA.

● Genetic engineering creates organisms with recombinant DNA.

Recombinant DNA: when DNA is combined from at least two organisms.

Techniques that 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. Therefore, DNA is not
recombined.