3 Quarter TOPICS

rd

• Genetic Engineering
• The Applications of Recombinant Deoxyribonucleic Acid (rDNA)
• The History of Life on Earth
• Mechanisms that Produced Change in Populations
• The Applications of Recombinant Deoxyribonucleic Acid (rDNA)
• Development of Evolutionary Thought
• Evidences of Evolution
• Evolutionary Relationships Among Organisms
• Linnaean System of Classification
• Biodiversity and Cladistics
4 Quarter TOPICS
th

• Reproduction in Plants and in Animals

• Animal and Plant Development
• Plants and Animals: Nutrient Procurement and Processing
• Gas Exchange in Plants and Animals
• Transport and Circulation
• Plant and Animal Regulation of Body Fluids
• Sensory and Motor Mechanisms
• Plant and Animal Organ Systems and their Functions (Plant and Animal Immune
System)
• Temperature Regulation in Plants and Animals
• Feedback Mechanism: Osmotic Pressure and Sugar Levels
What’s the difference?
What’s the difference?
What’s the difference?
What is Genetic Engineering?
The wide variety of
plant-based products
and fortified formulas
in the market today
has been made
possible by artificial
selection of desired
genes through genetic
engineering.
What is Genetic Engineering?
Genetic Engineering - a
gene modification
process wherein the
Deoxyribonucleic acid
(DNA) is transferred
from one organism to
another.
 What is Genetic Engineering?

Tobacco Plant glows in the dark (1986) Gene taken from firefly
Classical Breeding vs Genetic Engineering
Classical Breeding vs Genetic Engineering

Classical breeding focuses

on the mating of organisms
with desirable qualities or
traits while genetic
engineering involves
molecular techniques to
modify the traits of a target
organism.
Classical Breeding vs Genetic Engineering
• The modification of traits may involve
the introduction of new traits into an
organism or enhancement of a present
trait by increasing or disrupting the
expression of the desired gene.

• It has an application in the

pharmaceutical, industrial,
agricultural, medical, and other
industries.
GMO’s (Genetically Modified Organisms)
• Genetically modified
organisms have been subject
for public scrutiny whether it
is safe to use or ethically
accepted.

• These challenge the

researchers to prove the
significance of GMOs as a
breakthrough in science.
Processes of Genetic Engineering
• Stage 1. DNA Cleavage
• Stage 2. Production of Recombinant DNA
• Stage 3. Cloning
• Stage 4. Screening
Processes of Genetic Engineering
1. DNA Cleavage 

- a restriction
endonuclease is used
to cleave the source
DNA into a different
set of fragments. 
Processes of Genetic Engineering
2. Production of
Recombinant DNA 
- The fragments of
DNA are inserted into
plasmids that have
been cleaved with the
same restriction
endonuclease as the
source DNA.
Processes of Genetic Engineering
Plasmid insertion
Processes of Genetic Engineering
3. Cloning 
- The plasmids serve as
vectors that can introduce
the DNA fragments into
cells--- usually, but not
always bacteria. As each cell
produces, it forms a clone of
cells that all contain the
fragment-bearing vector.
Processes of Genetic Engineering
4. Screening 
- The clones containing a
specific DNA fragment
of interest are identified
from the clone library.
Methods of Introducing Plasmids to Host Organism
1. Biolistic
2. Plasmid insertion by Heat
Shock Treatment
3. Electroporation
Methods of Introducing Plasmids to Host Organism
1. Biolistic
- This technique uses a “gene
gun” to fire DNA-coated pellets
on plant tissues. Cells that are
able to survive and take up the
expression plasmid coated
pellets can acquire the ability to
express the designed protein.
Methods of Introducing Plasmids to Host Organism
2. Plasmid insertion by Heat
Shock Treatment
- is a process used to transfer
plasmid DNA into bacteria. The
target cells undergo a
pretreatment procedure to
increase the pore sizes of their
plasma membranes.
Methods of Introducing Plasmids to Host Organism
2. Plasmid insertion by Heat Shock Treatment
• The pretreatment using Calcium chloride makes the cells
“competent” for the introduction of the plasmid DNA, then the
cells are incubated with the desired plasmid at about 4°C for
about 30 minutes.
• During this time, the plasmids concentrate near the cells.
Afterward, a “Heat Shock” is done on the plasmid-cell solution by
incubating it at 42°C for 1 minute then back to 4°C for 2 minutes.
The rapid rise and drop of temperature increase and decrease the
pore sizes in the membrane, respectively.
Methods to Screen Recombinant Cells
Recombinant Cell
• These are cells that are made by
combining genetic material from
two different sources.

1. Selection of plasmid DNA containing cells

2.Selection of transformed cells with the desired gene
3. Polymerase Chain Reaction (PCR) detection of plasmid
DNA
Methods to Screen Recombinant Cells
1. Selection of plasmid DNA containing cells

• A selection marker within the inserted plasmid DNA sequence

allows the selection of “transformants”. Usually, an antibiotic
resistance gene is included in the plasmid DNA. This mechanism
allows only “transformed” cells to survive in the presence of the
antibiotic (e.g. ampicillin).
Methods to Screen Recombinant Cells
1. Selection of plasmid DNA containing cells
Methods to Screen Recombinant Cells
2. Selection of transformed cells with the desired gene

• The most general procedure for screening clone libraries to find

a particular gene is hybridization - the cloned genes form base-
pairs with complementary sequences on another nucleic acid.

• The complementary nucleic acid is called as probe because it is

used to probe for the presence of the gene of interest.
Methods to Screen Recombinant Cells
3. Polymerase Chain Reaction (PCR) detection of plasmid DNA

• the presence of the desired gene in the inserted plasmids may

also be confirmed through PCR amplification.

• PCR reactions specific for the desired gene may be done using
DNA from cells that would confirm the presence of the gene
within the samples.
Methods to Screen Recombinant Cells
3. Polymerase Chain Reaction (PCR) detection of plasmid DNA

• PCR reactions specific for plasmid sequences will confirm/identify the

type of plasmid used for the transformation through the following steps:

• Step 1. Denaturation.
• Step 2. Annealing of Primers.
• Step 3. Elongation.
Methods to Screen Recombinant Cells
3. Polymerase Chain Reaction (PCR) detection of plasmid DNA