Specific Learning

Outcome:

At the end of the

lesson, the learners
will be able to:
 Compare classical
breeding with modern
Specific Learning
genetic engineering
Outcome: techniques.

At the end of the

lesson, the learners
will be able to:
 Compare classical
breeding with modern
Specific Learning
genetic engineering
Outcome: techniques.

At the end of the

lesson, the learners
Enumerate the steps in
will be able to: Molecular Cloning.
 PRETEST
Direction: Read each
questions aloud. After,
choose the letter of the
best answer.
1. Which of the following would be
considered a transgenic organism?
A. Bacterium that has received genes
via conjugation
B. Human given a human insulin from
transgenic bacteria
C. Rat with a rabbit hemoglobin genes
D. Human treated with blood clotting
actors from bacteria
2. A scientist found out that a bacteria
infected by bacteriophages had
developed an ability to make an amino
acid that could not make before. These
new ability was probably a result of
A. Transformation
B. Natural selection
C. Conjugation
D. Transduction
For numbers 3-5, use the following choices:

A. Plasmid
B. Transduction
C. Conjugation

3. The transfer of bacterial gene

by a phage.
 A. Plasmid
B. Transduction
C. Conjugation
4. A circular DNA molecule that
is smaller and separate from the
bacterial chromosomes.
 A. Plasmid
B. Transduction
C. Conjugation
5. It refers to the union of cells
and the DNA transfers between
the two bacterial cells.
 ALL ABOUT GENETIC
ENGINEERING
• From the pass decades, humans look for
ways or methods in improving the quality of
their lives.
• They become immerse in improving the
quality of their domesticated plants and
animals.
• Scientists develop a technique that aims
to modify the genetic composition of an
organism to possess the desired traits or
phenotypes.
• Genetic engineering is like LEGO
blocks, where scientist build up DNA’s
until they form and create new
organisms.
 What is Classical Breeding?
• Involves mating of two members of a
species — each possesses one or more
different and desirable traits — to create
a hybrid individual possessing both
traits.
• Importantly, it does not involve any
direct manipulation of genetic material.
What is Genetic Engineering?
• A process that uses laboratory-
based technologies and molecular
techniques to alter the DNA makeup
of an organism.
• This may involve changing a single
base pair, deleting a region of DNA
or adding a new segment of DNA.
 What is Molecular Cloning?
• It involves separation of specific
gene or DNA fragments from a
donor cell, attaching it to small
carrier molecule called vector and
then replicating this recombinant
vector into a host cell.
Major Steps in Molecular Cloning
1. Isolation of donor DNA fragment or gene
2. Selection of suitable vector
3. Incorporation of donor DNA fragment into
the vector
4. Transformation of recombinant vector into
a suitable host cell
5. Isolation of recombinant host cell
 1. ISOLATION OF DONOR DNA
FRAGMENT OR GENE
• A donor DNA fragment should be
isolated.
• Restriction endonuclease enzyme: It has specific
restriction site for its action. The enzyme RE generates
a DNA fragment either with blunt end or with sticky
end.
• Reverse transcriptase enzyme: Synthesizes
complementary DNA strand of the desired gene using
its mRNA.
 2. SELECTION OF SUITABLE
VECTOR
• When donor DNA fragment is incorporated into a host
cell, it will not replicate because the isolated gene do
not have the capacity to replicated itself. So, before
introduction of donor fragment into host, a suitable
vector should be selected.
• Cloning vector is the DNA molecule capable of self-
replication inside the host cell. The main function of
cloning vector is to replicates the inserted DNA
fragment inside the host cell.
Characteristic of
Cloning Vector
1.It must be self-replicating inside host cell
2.It must possess restriction site for RE
enzymes
3.Introduction of donor DNA fragment must
not interfere with replication property of
the vector
4.It must possess some marker gene such
that it can be used for later identification
of recombinant cell.
 3. INCORPORATION OF DONOR DNA
FRAGMENT INTO THE VECTOR
• The plasmid vector is cut open by the same RE
enzyme used for isolation of donor DNA
fragment.
• The mixture of donor DNA fragment and
plasmid vector are mixed together.
• In the presence of DNA ligase, base pairing of
donor DNA fragment and plasmid vector occurs
forming recombinant vector in the mixture
 4. TRANSFORMATION OF RECOMBINANT
VECTOR INTO A SUITABLE HOST CELL
• The recombinant vector is transformed into
suitable host cell. Ex. bacterial cell.
• Some bacteria are naturally transformable, they
take up the recombinant vector automatically.
Ex: Bacillus, Haemophillus, Helicobacter pylori.
• Some other bacteria are not naturally competent,
in those bacteria recombinant vector are
incorporated by artificial method such as Ca++
ion treatment, electroporation etc.
 5. ISOLATION OF
RECOMBINANT HOST CELL
• The recombinant host cell is then grown in
culture media, but the culture may contain
colonies both recombinant cell and non-
recombinant cell.
• For isolation of recombinant cell from non-
recombinant cell, marker gene of plasmid
vector is employed.
 Activity No. 1
Part 1:
Direction: COPY & ANSWER in your
notebook. Match column A with the
correct description on Column B.
Simple WRITE the letter of your
answer on the space provided.
 Activity No. 1
Part 2:
Direction: COPY & ANSWER in your
notebook. Identify whether an
organism is a product of CLASSICAL
BREEDING or GENETIC
ENGINEERING.
 CLASSICAL BREEDING or GENETIC
ORGANISM
ENGINEERING

1. Plum trait in Apricot

2. Pigs with growth hormones coming from

cow

3. Pink flower

4. Pigs with human hemoglobin gene

5. Cows with BST to increase milk yield