RECOMBINANT DNA

TECHNOLOGY

PRIMING: Picture Analysis

1. What do you see in the
picture?
2. Are they the same with
the ordinary goldfish that
you know? Cite their
similarities and differences.

What is Recombinant
DNA Technology
(rDNA)?

Recombinant DNA Technology

molecules are formed by
laboratory methods of
genetic recombination
(such as molecular
cloning) to bring together
genetic material from
multiple sources, creating
sequences that would not
otherwise be found in
biological organisms.

 Cite of Cleavage A
specific sites at which
restriction enzyme will
cleave DNA.
Plasmid - plasmid is a
small DNA molecule within
a cell that is physically
separated from a
chromosomal DNA and
can replicate
independently.

Applications of rDNA

Recombinant Human Insulin

Almost completely
replaced insulin obtained
from animal sources (e.g.
pigs and cattle) for the
treatment of insulindependent diabetes.

Recombinant Human Growth

Hormone (HGH, somatotropin)

Administered to patients whose pituitary glands generate

insufficient quantities to support normal growth and
development.

Recombinant Blood Clotting Factor

VIII
It is a blood-clotting
protein which is
administered to patients
with forms of the bleeding
disorder hemophilia, who
are unable to produce
factor VIII in quantities
sufficient to support
normal blood coagulation.

Recombinant Hepatitis B Vaccine

Hepatitis B infection is controlled through the use of a

recombinant hepatitis B vaccine, which contains a form of the
hepatitis B virus surface antigen that is produced in yeast
cells.

Diagnosis of infection with HIV

Each of the three widely
used methods for
diagnosing HIV infection
has been developed using
recombinant DNA. The
antibody test uses a
recombinant HIV protein
to test for the presence of
antibodies that the body
has produced in response
to an HIV infection.

Golden rice

A recombinant variety of rice that has been engineered to

express the enzymes responsible for -carotene biosynthesis.
This variety of rice holds substantial promise for reducing the
incidence of vitamin A deficiency in the world's population.
Golden rice is not currently in use, pending the resolution of
regulatory issues.

Herbicide-resistant crops
Commercial varieties of
important agricultural crops
(including soy, maize/corn,
sorghum, canola, alfalfa and
cotton) have been developed
that incorporate a recombinant
gene that results in resistance to
the herbicide glyphosate (trade
name Roundup), and simplifies
weed control by glyphosate
application. These crops are in
common commercial use in
several countries.

Insect-resistant crops

Bacillus thuringeiensis is a bacterium that naturally produces

a protein (Bt toxin) with insecticidal properties. The bacterium
has been applied to crops as an insect-control strategy for
many years, and this practice has been widely adopted in
agriculture and gardening.

CLOSURE
APPLICATION

What are the benefits of recombinant DNA? Can we apply

those processes here in Bacoor?

EVALUATION

Give five examples of application of recombinant DNA.

CLONING

PRIMING
What do you know about the theory of creation?
How did God create man?

How would you feel if this happens

to you?

Cloning
Cloning in biotechnology
refers to processes used
to create copies of DNA
fragments (molecular
cloning), cells (cell
cloning), or organisms.

MAJOR STEPS IN
CLONING

1. Fragmentation
2. Ligation
3. Transfection
4. Screening/Selection

1. Fragmentation

breaking apart a strand of DNA.

2. Ligation
gluing together pieces of
DNA in a desired
sequence.

3. Transfection

inserting the newly formed pieces of DNA into cells.

4. Screening / Selection
selecting out the cells that
were successfully
transfected with the new
DNA

APPLICATION
Is cloning important to the
society?

Activity: Cloning?..
Instructions:
Read a part of the particle and answer the guide
questions.
Scientists Successfully Insert Woolly Mammoth
DNA Into Elephant Genome.*

*The Article was taken from iflscience.com

Scientists Successfully Insert Woolly Mammoth

DNA Into Elephant Genome.
In true "Jurassic Park" style,
scientists at Harvard University
have successfully managed to
insert genes from the woolly
mammoth into the genome of
an elephant. While this may
represent significant progress
in the field, lead researcher
George Church has reportedly
played down claims that the
work brings us closer to
recreating these iconic animals.

1.

What were the scientist from Harvard University able to

accomplish?

2.

Who is the lead researcher in the said article?

Scientists Successfully Insert Woolly Mammoth

DNA Into Elephant Genome.
Woolly mammoths (Mammuthus
primignius) may have appeared
more than 400,000 years ago
during the middle Pleistocene, but
they actually didnt die out all that
long ago. Alongside most other
large mammal species residing in
the Northern Hemisphere, they
disappeared from most of their
range across mainland Eurasia and
North America about 10,000 years
ago, but a small population of some
500-1,000 individuals survived on
Wrangel Island in the Arctic Ocean
for a further 6,000 years.

3. When did the Woolly mammoth first appear?

4. How many years has the Woolly Mammoth been extinct?

Scientists Successfully Insert Woolly Mammoth

DNA Into Elephant Genome.
Since many resided in frozen parts of the
world, when they died their bodies
sometimes became encased in
permafrost, which largely shielded them
from decomposition and the hungry
mouths of predators and scavengers.
This has meant that some remarkably
well-preserved specimens have been
recovered as the ice has thawed and
revealed their resting place, some of
which could be 40,000 years old. But
while they may look pretty intact, the
same cannot be said for their genomes
since DNA degrades over time, a process
that is accelerated by the presence of
microbes and water.

5. How is it possible for scientist to obtain Partial DNA from an

already extinct animal?

6. Why is it impossible for scientist to obtain full genome or

fully intact DNA from prehistoric organisms?

Scientists Successfully Insert Woolly Mammoth

DNA Into Elephant Genome.
Although scientists have managed to
find fragments of mammoth DNA
from frozen cells, which raised the
possibility they could be stitched
back together, they have so far failed
to find enough to perform cloning
experiments. While some scientists
have therefore ruled out using this
technique to bring mammoths back
from extinction, all hope may not be
lost as some think it is possible to
merge genes takenfrom preserved
specimens with those of their
closest living relativethe Asian
elephant.

7. What animal is said to be the closest living relative of the

Woolly Mammoth?

8. How are the scientist planning to create a clone of the said

animal?

Vectors used in
Genetic Engineering

Priming: Describe the illustration

below.

What is a vector?

Vector

an organism, typically a biting insect or tick, that transmits a

disease or parasite from one animal or plant to another.

Vector in Genetics
a
bacteriophage
or
plasmid that transfers
genetic material into a
cell,
or
from
one
bacterium to another.

Different Cloning
Vectors

What is this?

Bacterial Vectors
Bacteria harbor plasmids:
circular double strands of
DNA that are extrasomal
(Not part of the bacterial
chromosome).
These
plasmids
are
easily
manipulate
and
have
been
engineered
as
cloning
vectors
with
unique restriction sites for
insertion of foreign DNA

Bacterial Vectors

Bacterial plasmids can insert up to 10 kilobases of DNA (1

Kilobase is equals to 1000 nucleotides)

What is the
difference
between the
top and
bottom
picture?

Non-integrating Plasmid

Non-Integrating Plasmid
Non-integrating plasmids are the plasmids that doesnt
incorporate itself into the chromosomal DNA. When cell
division occurs, both daughter cells inherits both the
chromosomal DNA and the extrasomal Plasmid.

Integrating Plasmid

Integrating Plasmid
Integrating plasmid can incorporate itself into the
chromosomal DNA. The daughter cell produced contains
only the plasmid integraed DNA.

What are the things that affect the

success rate of DNA Cloning?
Some Plasmids or microbial host
include an addiction system of
postsegregational killing system
(PSK). One such example can be
found in E.Coli. The daughter
cell that retains a copy of the
plasmid
survive
while
the
daughter cell that fails to inherit
the plasmid dies or suffers a
reduced growth rate due to the
lingering poison from the parent
cell.

Screening for the successful DNA

Cloning

These plasmids contain a selectable marker, usually an

antibiotic resistance gene, which confer on the bacteria an
ability to survive and proliferate in a selective growth medium
containing the particular antibiotics. The bacteria are then
exposed to the selective media, and only cells containing the
plasmid may survive. In this way, the antibiotics act as a filter
to select only the bacteria containing the plasmid DNA.

USES OF PLASMIDS
Plasmid may also be used
for gene transfer into
human cells as potential
treatment
ingene
therapyso that it may
express the protein that is
lacking in the cells. Some
strategies
ofgene
therapyrequire
the
insertion
of
therapeuticgenesat pre-

DNA Vaccination

DNA vaccination is a technique for protecting an animal

against disease by injecting it with genetically engineered
DNA so cells directly produce an antigen, resulting in a
protective immunological response.

APPLICATION
If you were a famous geneticist, What are the problems
in Bacoor that you would like to solve using genetic
engineering using plasmids?

Assignment
1. Define bacteriophage.
2. Give and describe the different stages of
bacteriophage.

Bacteriophage

Priming 1: Describe what you see in

the picture below.

Priming: Arrange the jumbled letters

to form a new word.

CEPAGABIOTERH

What is a Bacteriophage?
It is a virus that infects
and replicates within a
bacterium.
A virus that infects a
bacterium is called a
bacteriophage.

What is a Bacteriophage?

Viral DNA can be engineered for use as a cloning vector.

The Viral DNA can be used to deliver a recombinant or

modified genes into a cell.

Life Cycle of a Bacteriophage

1. Phage attaches itself to
the surface of
bacterium.

Life Cycle of a Bacteriophage

2. Phage injects its DNA into the bacterium.

Life Cycle of a Bacteriophage

3. Phage Viral coat
separates from the cell
surface, leaving the DNA
inside the bacterium.

Life Cycle of a Bacteriophage

4. Phage DNA directs host cell to produce new viral proteins and
DNA.

Life Cycle of a Bacteriophage

5. Complete mature phages
assemble.

Life Cycle of a Bacteriophage

6. Cell wall bursts releasing 100 to 200 new phages.

Parts of a Bacteriophage

Insertion of DNA Bacteriophage