Senior High School

General Biology 2
Quarter 3 Week 1– Module 1:
Genetic Engineering and
Applications of Recombinant
DNA Technology

Photocredit.theweek.in
General Biology 2 – Grade 12
Quarter 3 – Module 1: Genetic Engineering and Applications of Recombinant
DNA Technology
Second Edition, 2021

Republic Act 8293, section 176 states that: No copyright shall subsist in
any work of the Government of the Philippines. However, prior approval of the
government agency or office wherein the work is created shall be necessary for
exploitation of such work for profit. Such agency or office may, among other things,
impose as a condition the payment of royalties.

Borrowed materials (i.e., songs, stories, poems, pictures, photos, brand

names, trademarks, etc.) included in this module are owned by their respective
copyright holders. Every effort has been exerted to locate and seek permission to use
these materials from their respective copyright owners. The publisher and authors
do not represent nor claim ownership over them.

Published by the Department of Education

Secretary: Leonor Magtolis Briones
Undersecretary: Diosdado M. San Antonio

Development Team of the Module

Compiler/ Contextualizer: EUSEBIA L. HERNANDEZ, MT1 Don Sergio
Osmeña Sr. Memorial National High School
Editors/Reviewers:
CELIA C. GEPITULAN - Principal 1, Tejero National High
School
JOCELYN C. BUTANAS - MT1, Talamban National High School
BONNIE JAMES SACLOLO - Teacher III, Cebu City National
Science High School
Language Editor
ROQUESA B. SABEJON-PSDS NORTH DISTRICT 7

Management Team:
Chairperson: DR. RHEA MAR A. ANGTUD - Schools Division Superintendent
DR. BERNADETTE A. SUSVILLA -Asst. Schools Division Superintendent
MRS. GRECIA F. BATALUNA Chief, Curriculum Instruction Division
MRS. VANESSA L. HARAYO- EPS, LRMDS
DR. RAYLENE S. MANAWATAO – EPS, SCIENCE

Department of Education – Schools Division of Cebu City, Region VII

Office Address: New Imus Road, Day-as, Cebu City, Philippines

Telefax: 032-2551516
E-mail Address: cebu.city@deped.gov.ph
Website: http://www.depedcebucity.com

2
 What I Need to Know

MODULE 1
Genetic Engineering and Applications of Recombinant DNA
Technology
__________________________________________________________________________________
_____
Quarter : Third Quarter
Content Standard : The learners demonstrate an understanding of recombinant
DNA.
Performance Standard: The learners should be able to make a research paper/case
study/poster on genetic diseases.
Learning Outcomes (Syllabus): Upon the completion of the given unit, the SHS
students are expected to explain the processes involved in
genetic engineering and how the technique is used recombinant
DNA technology.
Learning Competencies: Outline the processes involved in genetic engineering
(STEM_BIO11/12-IIIa-b-6)
Discuss the applications of recombinant DNA technology
(STEM_BIO11/12-IIIa-b-7)
Duration : 1 Week
Topics : Genetic Engineering
Recombinant DNA Technology
__________________________________________________________________________________

Hello STEM learners! In this module, you will explore the processes involved
in genetic engineering. This technology allows the manipulation of the organisms’
genetic components to make useful products that affect everything from agriculture
to criminal law to medical research. Furthermore, you will also familiarize some of
the products of recombinant DNA technology and consider some of the important
social and ethical issues that arise as biotechnology becomes more pervasive in our
lives. Spacing 1.15
After going through this module, you are expected to:
1. enumerate the steps in molecular cloning.
2. describe some methods to introduce DNA into cells.
3. explain the selection and screening of transformants/genetically modified
organisms (GMOs).
4. describe steps in Polymerase Chain Reaction (PCR) to amplify and detect a
gene of interest; and
5. enumerate some applications of recombinant DNA technology.

3
 What I Know

Directions: Write the letter of the correct answer on a separate sheet of paper.

1. Which of the following is NOT a product of genetic engineering?

A. macapuno C. Flavr Savr tomato
B. recombinant insulin D. Bacillus thuringensis (Bt) corn
2. Which of the following is the correct sequence of PCR amplification?
A. new strand elongation, primer annealing, undenatured template,
template denaturation
B. primer annealing, new strand elongation, undenatured template, template
denaturation
C. undenatured template, template denaturation, primer annealing,
new strand elongation
D. undenatured template, primer annealing, template denaturation, new
strand elongation
3. What happens to the organism (host cell) when a gene is inserted into it?
A. the organism might die
B. the organism might express the gene
C. there is no change in the organism’s traits
D. there is a big change in the organism’s traits
4. Which of the following is a characteristic of Bt corn?
A. pest-resistant C. herbicide-resistant
B. longer shelf-life D. enhanced nutrients
5. How does the desired gene obtain from the donor organism? The desired or
target gene is ____________________.
A. sealed by DNA ligase C. removed through a gene gun
B. exposed to heat shock D. cleaved by restriction enzymes
6. What enzyme cuts this DNA sequence 5’ ATGAGTTCCCCGAATGTC 3’?
A. EcoR1 B. Hind III C. Hind I D. Taq I
7. At what temperature does the denaturation of DNA double helix takes place?
A. 54°C B. 60°C C. 74°C D. 95°C
8. Which enzyme is used to join the bacterial plasmid and the inserted DNA
sequence?
A. DNA ligase C. DNA helicase
B. DNA gyrase D. DNA polymerase
9. Which of the following procedures occur after the desired gene has been
cleaved?
A. expression of the gene C. ligation of the gene of interest
B. insertion to the vector D. reproduction of the desired gene
10. What is the function of the antisense RNA inserted in Flavr Savr tomato
which has a delayed fruit ripening?
A. activator B. inhibitor C. regulator D. vector

4
11. Below are the steps in recombinant DNA technology. What is the correct
sequence of the given steps?
I. ligation (joining) of the gene of interest (eg. from an animal) with
the vector (cut bacterial plasmid)
II. cutting or cleavage of DNA by restriction enzymes (REs)
III. transfer of the recombinant plasmid into a host cell
IV. selection process to screen which cells contain the gene of interest
A. I, II, III, IV B. IV, III, II, I C. II, IV, I, III D. II, I, III, IV
12. Which of the following organisms can produce more copies of the
recombinant insulin gene in a shorter period?
A. bacteria B. mouse C. pig D. yeast
13. What happens during the denaturation step of a polymerase chain reaction?
A. The DNA nucleotides are broken apart.
B. The DNA is returned to its natural setting.
C. The base-pairing rules for DNA are reversed.
D. The double-stranded DNA is separated into two single strands of
DNA.
14. Which nutrient is highly present in genetically engineered golden rice?
A. collagen B. omega 3 C. Vitamin A D. beta-carotene
15. Which of the following steps happened during heat shock treatment?
A. The cell is exposed to an electrical shock.
B. The DNA particle is inserted into the cell through gamma rays.
C. DNA-coated pellets are fired to the plant tissues using a gene gun.
D. The cell is exposed to rapid rise and drop of temperature thereby
increasing pore size.

Genetic Engineering and Recombinant DNA Technology

What’s In
Review on Central Dogma of Molecular Biology: Unlocking the Code

Each organism has its genetic material (DNA) which must be replicated before
the cell undergoes cell division to ensure that the daughter cells contain the same
genetic material as the parent cell. During DNA replication, the cell requires different
proteins and enzymes (e.g., DNA polymerase, helicase, ligase) to carry out
complementary base pairing producing a new DNA strand for each unwound
template DNA. In transcription, the DNA strand is transcribed into messenger RNA
because the tRNA cannot read such code. During translation, the mRNA transcript
is read by the transfer RNA which brings an amino acid for each codon (3 letter-
bases). The code is now translated into proteins which are responsible for the
expression of the traits of the organism.
To further check if you still remember the concepts of the central dogma,
illustrate the steps during DNA replication, transcription, and translation using the
given DNA template. (Note: This activity simplifies the three processes hence only one

5
strand is given and the responsible enzymes and proteins for each step were not
included here).

REPLICATION
DNA template strand : ATG ACT CTA GAC TAA CTT ACC GAC TAG
Complementary strand : TAC _________________________________________________

TRANSCRIPTION
mRNA transcript : AUG__________________________________________________

TRANSLATION (tRNA brings a specific amino acid for each mRNA

codon kindly refer the genetic code chart below as your
guide: read the mRNA not the tRNA)
tRNA/anticodon : UAC__________________________________________________
Amino acid/proteins : met-__________________________________________________

Genetic code chart (https://creativecommons.org/licenses)

Guide Questions

1. What happens during replication?

__________________________________________________________________________
2. Why is it important to transcribe the DNA into mRNA before translation?
__________________________________________________________________________
3. What is the function of tRNA in translation?
___________________________________________________________________________
4. Explain the importance of DNA in the expression of traits among organisms.
___________________________________________________________________________
5. How does the insertion of a desirable gene into the target organism affect its
characteristics?
___________________________________________________________________________

6
 What’s New

Desirable Traits
Identify how each of the traits was introduced or developed (i.e., classical breeding
or genetic engineering) in the given organisms below. Then, answer the guide
questions that follow. Write your answers on a separate sheet of paper.

1. _____________________________ 2._______________________________

3._____________________________ 4. ____________________________
Figure 1. GMOs. a.) Wagyu beef (https://caneridegecattle.com) b.) macapuno
(https://foodrecap.net) c. Bt corn (https://philstar.com)
d. GM soybeans (https://bearmarketreview.wordpress.com)

Guide Questions
5.What is genetic engineering?
________________________________________________________________________

6.What are genetically modified organisms (GMOs)?

_______________________________________________________________________

7.What is the advantage of genetic engineering/recombinant DNA technology

over classical breeding in terms of trait enhancement among organisms?
___________________________________________________________________________

What Is It

Humans have successfully domesticated selected plants and animals

particularly in choosing the desired traits. Traits that were considered valuable (i.e.
high fruit yield; high milk production, etc.) were sought out and propagated. The

7
processes involved may include classical breeding practices such as controlled
pollination of plants and the mating of animals with desired traits. In today’s modern
science, molecular biology techniques are being employed in the insertion and
expression of proteins in different organisms for various purposes. Genetic
engineering involves the use of molecular techniques to modify the traits of the target
organism. The modification of traits may involve:

A. Introduction of new traits into an organism

• A new trait may develop after the gene responsible for the trait is
inserted. Ex. glowing mice- inserted with the bioluminescent gene from
jellyfish
B. Enhancement of a present trait by increasing the expression of the
desired gene
• A present trait that is slightly expressed may be enhanced after the
insertion of an enhancer gene. Ex. golden rice-naturally rice can
synthesize beta-carotene in low concentration but may be enhanced
after inserted with a gene from the plants with high beta-carotene (e.g.,
carrots) which is a precursor of Vitamin A synthesis. Golden rice helps
in solving Vitamin A deficiency in children. Insufficient Vit. A leads to
blindness.
C. Enhancement of a present trait by disrupting the inhibition of the
desired genes’ expression
• The expression of the trait may be disrupted by introducing an inhibitor
(stops/ slows down the expression of the trait) like the ripening
processes in tomatoes. Ex. In Flavr-Savr tomatoes- an inhibitor (i.e.,
antisense RNA) disrupts the expression of the enzyme that causes
degradation of pectin in cell walls, thereby delaying the softening of the
fruit thus, increasing the shelf life.

Figure 2. Genetically modified organisms: Golden rice (www.irri.org) and Flavr Savr
tomato (http://ucce.ucdavis.edu/)

General Outline of Recombinant DNA Technology

I. Cutting or cleavage of target DNA sequence by restriction enzymes (REs) or
restriction endonucleases
➢ The REs cut the DNA molecules at a limited number of specific
locations. It can recognize the restriction site and cuts both DNA
strands at precise points with a sticky end (single-stranded end).
These short extensions can form hydrogen-bonded base pairs with
complementary sticky ends on any other DNA molecules cut

8
with the same enzyme (refer to Figure 2). Here are the common examples of
REs:
a. EcoR1(from Escherichia coli) cuts at
5’ GAATTC 3’
3’ CTTAAG 5’
b. BamH1(from Bacillus amyloliquefaciens) cuts at
5’ GGATTC 3’
3’ CCTAAG 5’

➢
Figure 3. Steps in recombinant DNA technology
(https://creativecommons.org/licenses

9
 II. Selection of an appropriate vector or vehicle which would
propagate the recombinant DNA
➢ The most common vector used in propagating the desired gene (e.g.
circular plasmid in bacteria with a foreign gene of interest) is the
bacteria and yeast. These organisms are widely used because they
can reproduce exponentially in a short period.
III. Ligation of the gene of interest with the vector
➢ The ligation of the two DNA strands (e.g., from animal and cut
bacterial plasmid) is permanently sealed by an enzyme DNA ligase.
This catalyzes the formation of covalent bonds of the sugar-
phosphate backbones that results in a stable recombinant DNA
molecule (e.g., bacterial plasmid).
IV. Transfer of the recombinant plasmid into a host cell
➢ The cell carries out replication to make huge copies of the
recombined plasmid. The recombinant plasmid can be transferred
through biolistic, heat shock treatment, or electroporation
depending on the recipient organism.
V. Selection process to screen the cells containing the gene of interest
➢ Cells containing the gene will be screened through selection
markers like antibiotic gene, fluorescent gene, or through a
polymerase chain reaction of the plasmids.
VI. Sequencing of the gene to find out the primary structure of the protein
➢ Once the cells with the gene have been identified, DNA sequencing
is employed to separate pieces of DNA that differ in length by only
one base. The most common technique is through gel-
electrophoresis wherein the DNA to be sequenced is placed at one
end of a gel - a slab of a gelatin-like substance.

Methods of plasmid insertion to the target organism

1. Biolistic. In this technique, a “gene gun” is used to fire DNA-coated pellets on

plant tissues (Figure 4). Cells that survive the bombardment and can take up the
expression plasmid-coated pellets and acquire the ability to express the designed
protein.
2. Plasmid insertion by Heat Shock Treatment. This technique is used to
transfer plasmid DNA into the bacteria. The cell is exposed to a “heat shock”
(rapid rise and drop of temperature) which increases and decreases the pore sizes
in the membrane. The plasmid DNA near the membrane surface is taken into
the cells by this process. The cells that took up the plasmids acquire new traits
and are said to be “transformed”.
3. Electroporation. This technique follows a similar methodology as Heat Shock
Treatment, but the expansion of the membrane pores is done through an electric
“shock”. This method is commonly used for the insertion of genes into
mammalian cells.

Figure 5 in the next page shows the differences in heat shock and
electroporation technique of plasmid insertion to the host cell.

10
Figure 4. Biolistic method in plants. (https://www.intechopen.com/chapters/30876)

Chemical transformation

Incubation Heat shock Recovery

e.g.,42oC, 30s

Chemically
Competent cell

Electroporation

Incubation Electric shock Recovery

Incompetent cell

Figure 5. Heat shock vs. Electroporation of Plasmid Insertion

(https://www.thermofisher.com)

11
Methods to screen recombinant cells

1. Selection of plasmid DNA containing cells

A selection marker (e.g., AMP ampicillin resistance gene) is included in the
plasmid DNA (refer to Figure 3). This allows only “transformed” cells to survive in the
presence of the antibiotic (e.g., ampicillin). Plating the plasmid-cell solution on
antibiotic-containing media will select for these “transformants” and only allow
plasmid-containing cells to grow and propagate into colonies.

2. Selection of transformed cells with the desired gene

Certain inserted genes within the plasmids provide visible proof of their
presence. Some inserted genes also produce colored (e.g., chromogenic proteins) or
fluorescent products that label the colonies/cells with the inserted gene.
3. Polymerase Chain Reaction (PCR) detection of plasmid DNA

The presence of the desired gene in the inserted plasmids may be confirmed
using PCR amplification. The DNA from the cells will be used in producing large
copies of the gene which will confirm the presence of the gene in the samples. (A
detailed discussion on the steps of PCR is found in the succeeding pages). Detection
of the presence of the SARSCOV 2 or the COVID-19 virus in the swab or saliva of the
patient use RT-PCR or Reverse Transcriptase-Polymerase Chain Reaction. However,
before the DNA amplification of the said virus, its genetic material must undergo
reverse transcription (meaning from RNA to DNA using the enzyme reverse
transcriptase) because it is an RNA virus. This is done to obtain the complementary
DNA which can undergo base pairing of the PCR primer. Running the DNA through
PCR cycles produces multiple copies of the gene which confirms the presence of the
virus in the sample hence the patient is considered positive.

Polymerase Chain Reaction (PCR)

PCR amplification is an in-vitro (occur outside a living organism) method that

simulates DNA replication in vivo (occur inside a living organism). It utilizes a
thermostable (heat-resistant) DNA polymerase that builds single-stranded DNA
strands to unwind DNA templates. PCR uses repeated cycles of incubation at
different temperatures to promote the unwinding of the DNA template (~95°C); the
annealing of a primer (a ~20bp oligonucleotide sequence (recall RNA primers in DNA
replication) into the single-stranded DNA (ssDNA) template strand (~54 - 60°C); and
the extension of the generated ssDNA strand through the binding of complementary
bases to the template strand (~72° C). The thermostability of the polymerase allows
it to survive the repeated cycles of denaturation, annealing, and extension with little
loss of enzyme function. Each cycle of PCR doubles the amount of the target
sequence. A typical PCR experiment uses about 35 cycles of amplification. This
increases the original amount of the target sequence by 235 (i.e. ~34 billion) times.
Unlike DNA replication in vivo, PCR reactions do not use too many helper enzymes
such as helicases and gyrases to help denature and stabilize the template DNA
strands. The cyclic heating of the samples is meant to provide the physical separation
of the template DNA strands through heat denaturation of the inter-strand H-bonds.

12
 The figure below summarizes the steps of Polymerase Chain Reaction. Observe
what happens to the DNA and how many strands are produced after one cycle.

Figure 6. Steps in Polymerase Chain Reaction

(https://creativecommons.org/licenses)

Steps in Polymerase Chain Reaction

Step 0: Undenatured Template at Temp ~ 54 °C
The complementary sequences of template double-stranded (ds) DNA
strands are held together by H-bonds.
Step 1: Template denaturation at Temp ~ 95 °C
The H-bonds between DNA complementary sequences are broken producing
single-stranded (ss) DNA strands that serve as the template.
Step 2: Primer Annealing at Temp ~ 54 °C (dependent on primer melting
temperature)
H-bonds are formed between complementary sequences on the primers and
the target sequences.
Step 3: New DNA strand elongation at Temp ~ 72 °C
The two new dsDNA strands are formed by the elongation of the generated
ssDNA and the H-bonds between the complementary sequences on these new
strands and their templates. Each of the new dsDNA strands is made up of
one old strand from the original template, and one new strand that was

13
 generated as a reverse complement of the template. This is called
semiconservative replication of the sequence.
Step 4: Repeat step 1 to 3 for N number of cycles (N is usually 35)
Each new DNA undergoes another cycle of PCR usually until 35 cycles
producing multiple copies of DNA in each cycle.

PCR may be used to detect the presence of the desired gene in an organism.
Depending on the primer design, the expected product may represent only a specific
region of the gene or the entire gene itself. The first case is useful for the detection of
the gene, or the detection of organisms with that specific gene within a sample. The
second case is useful for the amplification of the entire gene for eventual expression
in other organisms. The direct amplification/copying of a full gene is part of the
process for “cloning” that gene. Some genes provide economically, and industrially
important products (e.g. insulin-coding genes; genes for collagen degradation). In
some cases, scientists would want to put these genes into organisms for the
expression of their products. One example would be the insertion of an insulin coding
gene from the human genome into bacteria. This allows the “transformed” bacteria
to now produce human insulin as a product. This technology promises a lot of
applications in different fields which improves the lives of humanity.
Applications of Recombinant DNA technology
Recombinant DNA technology is the joining together of DNA molecules from
two different species. The recombined DNA molecule is inserted into a host organism
to produce new genetic combinations (new or improved traits) that are of value to
science, medicine, agriculture, and industry. This is the basis for the development of
genetically modified organisms (GMOs). GMOs are organisms whose genome has
been engineered in the laboratory to favor the expression of desired physiological
traits or the generation of desired biological products. The table below shows
examples of modified traits using cloned genes and their applications:
Modified Gene modification Recipient Application (field)
trait Organism
1. Insulin Insertion of human Bacteria Medicine: human insulin
production insulin gene for diabetic patients
2. Human Insertion of human Bacteria Medicine: mass
Growth growth hormone gene production of HGH for
Hormone children with dwarfism
3.Pest Insertion of Bt (Bacillus Corn (Bt Agriculture: increase
resistance thuringensis)-toxin gene that corn), crop production due to
kills only the corn borer Eggplant less pest infestation
larvae (but is harmless to (Bt-eggplant)
humans and other animals).
4.Delayed Disruption of a gene for a Tomato Agriculture: allow fruits
ripening ripening enzyme (Flavr-Savr) to survive longer
(polygalacturonase) transport time
5.Chymosin Insertion of a gene for Bacteria Industry: production of
production chymosin chymosin as rennet
(enzyme for coagulation
of milk- in cheese
making) substitute

14
 Most of the genetically modified organisms are plants. Some of these are
drought, salinity, frost, and herbicide-resistant plants that promote high crop
production which is of great help to the growing human population of the world.
There are also transgenic animals that serve as “pharmaceutical factories” which
helps in the mass production of proteins. For example, a transgene for a human
blood protein such as antithrombin, which prevents blood clots, can be inserted into
the genome of a goat in such a way that the transgene’s product is secreted in the
animal’s milk. However, some genetically engineered microbes are important in
mining (cleaning up highly toxic wastes), oil spills, and wastewater treatment.

Figure 7. Recombinant Products. Human Insulin injection (https://www.sedico.net)

and HGH (https://dfwantiagingwellness.com)
Genetic engineering promises a lot of benefits to humans especially for the
improvement of agricultural productivity and food quality. However, as responsible
consumers, we must remember that these products might bring potential harm to
humans, animals, and the environment as well. That is why consumption of GM
plants and animals and even the use of genetically engineered pharmaceutical
products pose a lot of ethical issues until now.

What’s More

Detecting SARS CoV2 through RT-PCR

The Severe Acute Respiratory Syndrome (SARS) Corona Virus 2 is an RNA

virus that is the causative agent of the global pandemic Coronavirus disease 2019.
This virus has a single-stranded RNA which can be found in respiratory droplets
from the patient and considered only living once infected a host (e.g. humans) which
leads to fever, shortness of breath, and other symptoms of the said disease. Once the
person has the symptoms, a swab test must be done to obtain a sample that may
contain the virus. This is done by using primers that are specific for complementary
DNA (cDNA) sequences that correspond to the SARS CoV 2 virus. If PCR amplification
indicates the presence of the said gene sequence, then the patient likely is considered
positive. Specifically, the virus can be detected through RT-PCR. In this technique,
the RNA population is converted to cDNA by reverse transcription (RT), and then the
cDNA is amplified by the polymerase chain reaction. The cDNA amplification step
provides opportunities to further study the original RNA species, even when they are
limited in amount or expressed in low abundance.

15
 Assuming that reverse transcription has been carried out and you already
obtained the cDNA, simulate the steps in PCR of the SARSCov2 on a piece of paper
for at least 3 cycles.

Materials
1 piece of double-stranded paper ‘template’ DNA, primers, nucleotides, scissors,
sticky tape

Procedure

Before simulating PCR prepare the following: (Remove the detachable template found
on the last page of this module; you may photocopy for backup copies).

a. Copy of DNA template and primers.

DNA template: 1st and 3rd lines are the first strands while the 2nd and 4th lines are
the second strands and when connected it should have the sequence as shown below:

A C C G A T C T C A G A T G A

T G G C T A G A G T C T A C T

T A G A C C G A T A G G A A G

A T C T G G C T A T C C T T C

ACCGATCTCAGATGATAGACCGATAGGAAG
TGGCTAGAGTCTACTATCTGGCT ATCCTTC

Primers: cut these to have 2 sets (see detachable sheet)

C C G A T A T C C T

C C G A T A T C C T

C C G A T A T C C T

C C G A T A T C C T

b. Individual cut-outs of the nucleotides (A, T, C, G).

A A A A A A A A A A A A A A

T T T T T T T T T T T T T T

C C C C C C C C C C C C C C

G G G G G G G G G G G G G G

Cycle 1

1.Use scissors to cut the double-stranded template DNA in half. (Caution: Be careful
in using the scissors).

16
(This represents the high-temperature denaturation step (95°C) in PCR, which breaks
the hydrogen bonds between the two DNA strands).
2. Match the primers next to their complementary sequences on the single-stranded
template DNA and stick them in place with tape.
(This represents primer annealing to the template DNA at the low-temperature
annealing step (55°C) in PCR. The primer annealing sites on the template DNA were in
bold to assist students in the first cycle of the activity – in future cycles, they will need
to find the annealing sites themselves).
3.Extend a new sequence from the primers using tape to attach each new nucleotide
to the previous nucleotide and the template strand. (Lay down a long strip of tape
first and then stick the new nucleotides onto it in turn).
(This represents the action of DNA polymerase in the 72°C extension step of PCR, which
creates a new strand of DNA starting at the 3’ end of each primer, incorporating new
nucleotides complementary to the template strand. The product is a double-stranded
section of DNA with one strand made up of the original template DNA and the other
made of newly synthesized DNA).

1. After cycle 1, you produce 2 new DNA strands. Repeat steps 1-3 to simulate
cycle 2 for each new DNA strand. At the end of the process, you will produce
4 new DNA strands. PCR usually reaches 35 cycles which produce multiple
copies of DNA.
2. Submit your PCR product to your teacher.

Guide Questions

1. How many DNA strands will be produced after 10 cycles?

__________________________________________________________________________
2. Explain the importance of Polymerase Chain Reaction in SARSCov2
detection? ________________________________________________________________
__________________________________________________________________________

What I Have Learned

To check your understanding of the concepts about genetic engineering and

its applications, write a short sentence or phrase on the given terms below.

Important term/concept Description

Genetic engineering
Recombinant DNA technology
Genetically modified organism
Restriction enzyme
Vector
Polymerase Chain Reaction

➢ Now it’s time to apply what you learned. The next activity tests your
creativity and curiosity. Good luck!

17
 What I Can Do

Designer Genes
Objectives: Produce a hypothetical genetically modified organism

Procedure

1. Identify a special trait to produce hypothetically modified or genetically

engineered plants and animals that can be used for health, industry,
agriculture, and the protection of the environment and its source organism.
2. Identify the source of the trait and your target organism.
3. Illustrate the steps to produce your designer organism (hypothetical
genetically modified organism) following the general steps in genetic
engineering.
4. Discuss the potential benefits and harm or ethical issues that your
recombinant organism brought to society.
5. Write your output in a short bond paper.
Criteria for Rating the Output

1. Originality of the study (i.e. Has anyone done studies of this type before?)
20 pts
2. Feasibility of the study (How possible is the proposed modification? Can
the target organism support the proposed trait?) 20 pts

3. Potential Applications of the new organism (What benefits would the

recombinant organism provide to society?) 20 pts

Assessment

Directions. Read each question comprehensively and write the letter of the correct
answer on a separate sheet of paper.
1.What is the function of gel electrophoresis?
A. cuts DNA C. recombines DNA
B. extracts DNA D. separates DNA fragments
2.Which of the following steps is NOT essential in producing recombinant
DNA?
A. amplify the entire DNA sequence
B. use a restriction enzyme to form sticky ends in DNA
C. splice the target DNA fragment with the bacterial plasmid
D. read the DNA sequence of the piece of DNA to be cut and spliced
3.Which of the following is often used as a genetic marker?
A. foreign gene
B. gene for antibiotic resistance
C. nucleotide labeled with a fluorescent dye
D. DNA sequence that serves as a bacterial origin of replication

18
4.What is the advantage of using transgenic bacteria to produce human proteins?
It can _____________________________________________.
A. be used to make plastics
B. produce human proteins in large amounts
C. last longer than those produced by humans
D. work better than those produced by humans
5.Which of the following is an advantage of producing transgenic plants?
A. producing clones C. studying human genes
B. using more pesticides D. increasing the food supply
6. What would be the effect on the PCR reaction if any of the following
circumstances arise: 1) there are no primers in the reaction, 2) there are no
dNTPs in the reaction, 3) there is no Taq polymerase in the reaction?
A. PCR would proceed normally.
B. The PCR reaction will not begin.
C. The reaction will cease after a few cycles.
D. Non-specific PCR of random templates will occur.
7.Which of the following steps should be done first to produce transgenic
bacteria that make human growth hormone? The first step to be done to
produce HGH is to __________________________________.
A. extract the insulin from the bacterial culture
B. insert the human insulin gene into a plasmid
C. transform bacteria with the recombinant plasmid
D. cut out the insulin gene from human DNA using restriction
enzymes
8. Which of the following is the first and the most important step in the
polymerase chain reaction?
A. extension C. dehydration
B. annealing D. denaturation
9.What is the enzymatic function of restriction enzymes?
A. join nucleotides during replication
B. cleave nucleotides at specific sites
C. join nucleotides during transcription
D. add new nucleotides to the growing strand of DNA
10. How many DNA duplexes are obtained from one DNA duplex after 4 cycles of
PCR?
A. 4 B.8 C.16 D. 32
11. What is used as primers in a polymerase chain reaction?
A. single-stranded RNA oligonucleotide
B. single-stranded DNA oligonucleotide
C. single-stranded DNA oligonucleotide
D. double-stranded RNA oligonucleotide
12. At what temperature does annealing of DNA and primer take place?
A. 42°C B.54°C C. 74°C D.95°C
13. What is used as a template during reverse transcription-polymerase chain
reaction (RT-PCR)?
A. DNA B. mRNA C. RNA D. tRNA
14. Which of the following animals is used as “pharmaceutical factories” for
antithrombin?
A. cow B. dog C. goat D. pig

19
15.What is the most logical sequence of steps for splicing foreign DNA into a
plasmid and inserting the plasmid into a bacterium?
I. Extract plasmid DNA from bacterial cells.
II. Cut the plasmid and target DNA using restriction enzymes.
III. Splice or join the bacterial plasmid and the desired DNA fragment.
IV. Transform bacteria with recombinant DNA molecule in a culture
medium.
A.I, II, IV, III B. I, II, III, IV C. I, III, IV, II D.II, IV, I, III

References

Offline sources

Reece, J.B; Urry, L.A; Cain, M.L; Wasserman, S.A; Minorsky, P.V; and Jackson, R.B.
(2014). Campbell Biology 10th. San Francisco (CA): Pearson Benjamin
Cummings.
Teaching Guide in General Biology 1. Department of Education. 2016
Online sources
https://creativecommons.org/licenses
https://caneridegecattle.com
https://foodrecap.net
https://philstar.com
https://bearmarketreview.wordpress.com
www.irri.org
http://ucce.ucdavis.edu
https://www.thermofisher.com
https://www.sedico.net
https://dfwantiagingwellness.com

20
 Answer Key

What is in
Suggested answers
1. The DNA strand undergoes complementary base pairing producing new strand.
2. Transcription must occur prior to translation because the tRNA cannot read the
DNA code (notice that in RNA Adenine pairs with uracil not thymine which is
only found in DNA).
3. tRNA brings amino acid for a specific codon (triplet nitrogen bases). This amino
acid will from a polypeptide to form the proteins which will be expressed by the
organism.
4. The genetic material of the organism is contained in the DNA. This DNA codes
for the gene responsible for the characteristics of the organism.
5. Insertion of a desired gene may enhance the characteristics of the organism.

What’s new
Suggested answers
1. Classical breeding 2. Classical breeding
3. Genetic engineering 4. Genetic engineering
5.Genetic engineering, also called recombinant DNA technology, involves the group
of techniques used to cut up and join genetic material, especially DNA from different
biological species, and to introduce the resulting hybrid DNA into an organism in
order to form new combinations of heritable genetic material.
6. Genetically Modified Organisms (GMOs) are organisms with genetically altered
genome to favor the expression of the desired traits.
7. Genetic engineering can insert genetic material from any life form into any other
whereas, classical breeding generally can only work within a species, or at most,
within closely related genera, as when they do wide crosses

What’s more
1. 1024
2. Scientists amplify a specific part of the transcribed viral DNA hundreds of
thousands of times. Amplification is important so that, instead of trying to spot a
minuscule amount of the virus among millions of strands of genetic information,
scientists have a large enough quantity of the target sections of viral DNA to
accurately confirm that the virus is present.

Answers for What I have learned and What I can do may vary

21
 DETACHABLE SHEET FOR WHAT’S MORE

DNA template

A C C G A T C T C A G A T G A

T G G C T A G A G T C T A C T

T A G A C C G A T A G G A A G

A T C T G G C T A T C C T T C

RNA primers

C C G A T A T C C T

C C G A T A T C C T

C C G A T A T C C T

C C G A T A T C C T

C C G A T A T C C T

C C G A T A T C C T

C C G A T A T C C T

C C G A T A T C C T

Nucleotides

A A A A A A A A A A A A A A

T T T T T T T T T T T T T T

C C C C C C C C C C C C C C

G G G G G G G G G G G G G G

A A A A A A A A A A A A A A

T T T T T T T T T T T T T T

C C C C C C C C C C C C C C

G G G G G G G G G G G G G G

22
 For inquiries or feedback, please write or call:

Department of Education - Bureau of Learning Resources (DepEd-

BLR)

Ground Floor, Bonifacio Bldg., DepEd Complex

Meralco Avenue, Pasig City, Philippines 1600

Telefax: (632) 8634-1072; 8634-1054; 8631-4985

Email Address: blr.lrqad@deped.gov.ph * blr.lrpd@deped.gov.ph

23