8

Biotechnology
Quarter 3 – Module 1:
Tools Used in Genetic Engineering

DIVISION OF ANGELES CITY

Biotechnology – Grade 8
Alternative Delivery Mode
Quarter 3 – Module 1-2: Tools Used in Genetic Engineering
First Edition, 2021

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Editors: Sherilyne L. Reyes, Jennifer Praza, Edgardo D. Cortez,
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 8
Biotechnology
Quarter 3 – Module 1:
Tools Used in Genetic
Engineering
Introductory Message
This Self-Learning Module (SLM) is prepared so that you, our dear learners,
can continue your studies and learn while at home. Activities, questions, directions,
exercises, and discussions are carefully stated for you to understand each lesson.

Each SLM is composed of different parts. Each part shall guide you step-
bystep as you discover and understand the lesson prepared for you.

Pre-tests are provided to measure your prior knowledge on lessons in each

SLM. This will tell you if you need to proceed on completing this module or if you
need to ask your facilitator or your teacher’s assistance for better understanding of
the lesson. At the end of each module, you need to answer the post-test to selfcheck
your learning. Answer keys are provided for each activity and test. We trust that you
will be honest in using these.

In addition to the material in the main text, Notes to the Teacher are also
provided to our facilitators and parents for strategies and reminders on how they can
best help you on your home-based learning.

Please use this module with care. Do not put unnecessary marks on any part
of this SLM. Use a separate sheet of paper in answering the exercises and tests. And
read the instructions carefully before performing each task.

If you have any questions in using this SLM or any difficulty in answering the
tasks in this module, do not hesitate to consult your teacher or facilitator.

Thank you.
 What I Need to Know

This module was designed and written with you in mind. It is

here to help you familiarize the different tools used in genetic engineering. The scope
of this module permits it to be used in many different learning situations. The
language used recognizes the diverse vocabulary level of students. The lessons are
arranged to follow the standard sequence of the course. But the order in which you
read them can be changed to correspond with the textbook you are now using.

The module is summated into one lesson, namely:

● Lesson 1 – Tools Used in Genetic Engineering

After going through this module, you are expected to describe the different tools used
in genetic engineering.
 What I Know

Directions: Read each question carefully. Choose the letter of the

correct answer.

1. The term is defined as the basic physical and functional unit of heredity.
a. alleles c. genes
b. deoxyribonucleic acid d. ribonucleic acid
2. It is a branch of science that is concerned how various traits are inherited.
a. biological engineering c. heredity
b. genetics d. pathology
3. The process by which recombinant DNA technology is utilized to alter the genetic
makeup of an organism.
a. biological engineering c. genetic engineering
b. biological modification d. genetic modification
4. When did Hindll isolate the first restriction enzyme?
a. 1960 c. 1990
b. 1980 d. 1970
5. It is the first organism used in DNA technology experiments.
a. E. coli c. S. cerevisiae
b. E. faecalis d. S. typhimurium
6. This is a laboratory technique used to amplify a single copy or a few copies of a
segment of DNA across several orders of magnitude, generating thousands to
millions of copies of a particular DNA sequence.
a. Agarose Gel Electrophoresis c. Polymerase Chain Reaction
b. Molecular Scissor d. Restriction Endonuclease
7. The enzymes that make one incision on each of the two strands of DNA at specific
locations based on the nucleotide sequence are known as ______________.
a. DNA Polymerase I c. Primase
b. Helicase d. Restriction Endonuclease
8. He discovered the Polymerase Chain Reaction.
a. Hamilton Smith c. Kary Mullis
b. Hindll d. Werner Abner
9. It is a branch of biotechnology that deals with the study and investigation of
genomic information from trace evidence found at crime scenes. Agricultural
a. biotechnology c. Industrial biotechnology
b. Forensics biotechnology d. Medical biotechnology
10. It is a technique commonly used in laboratories to separate charged molecules
like DNA, RNA and proteins according to their size.
a. Gel Electrophoresis c. Polymerase Chain Reaction
b. Molecular Scissor d. Selection of Prokaryotic Host
Lesson
The Different Tools Used in
1 Genetic Engineering
What’s In

Our previous module learned that mutation could result from DNA copying
mistakes during cell division, exposure to ionizing radiation, exposure to chemicals
called mutagens, or infection by viruses. The majority of mutations have neither
negative nor positive effects on the organism in which they occur. These mutations
are called neutral mutations. While there is a mutation with positive effects, a
mutation can develop resistance to some diseases like atherosclerosis. These
mutations are called beneficial mutations. Lastly, a harmful mutation can cause
genetic disorders and diseases such as cancer and cystic fibrosis.

ACTIVITY 1
Direction: Match the types of mutation to their genetic codes and write the letter of
the correct answer before each number.

TYPES OF MUTATION GENETIC

CODES

1. Duplication A.

2. Inversion B.

3. Normal Sequence C.

4. Deletion D.
 5. Insertion E.

ACTIVITY 2
Direction: Fill in the diagram with the different types of mutation.
(Note to the learners: You may draw the illustrated diagram on your answer sheet.)

Mutation
 What’s New
Direction: Match each tool used in genetic engineering to its
corresponding description. Choose the letter of the correct answer.

Answer Genetic Tools Description

1. Molecular Scissor
a. It multiplies the DNA
exponentially for each of the 25 to
75 cycles.
2. Polymerase Chain
Reaction b. It is the enzymes that cut
DNA at specific locations based on
the nucleotide sequence.

3. Gel Electrophoresis
c. It monitors the changes in
protein content in body fluids

d. The utilization of multi-cell

4. Polymerases organisms to produce human
proteins since these hosts with
complex structures are more
suitable for synthesizing complex
proteins.
5. Eukaryotic Host

e. The groups of enzymes that

catalyze the synthesis of nucleic
acid molecules

What do you think are these tools for in genetic engineering?

What is It
You may have heard about genetic engineering in newspapers,
TV shows, and the Internet. Sci-fi movies like X-Men and Splice depict individuals
with enhanced genetic modifications that give them special abilities.

So, what exactly is Genetic Engineering? Genetic engineering involves

manipulating genetic material (DNA) to achieve the desired goal in a predetermined
way. In history, one of genetic engineering’s breakthroughs was the successful
discovery of CRISPR/Cas9, which was used nowadays as an editing tool to modify
genomes—a trailblazing development for medicinal purposes curing genetic disorders
in the near future. Furthermore, genetic engineering also made cloning possible as it
successfully cloned a mammal from an embryo cell, a sheep named “Dolly,” and Ian
Wilmut and his colleagues executed the said cloning.

However, what are the standard tools used in genetic engineering? Moreover,
how does genetic engineering help in the detection of viruses?

Figure 1: Genetically Modified Organisms and Tools Used in Genetic Engineering.

Genetic Engineering Defined

The term genetic Engineering is initially referred to various techniques used for the
modification or alteration of organisms through the processes of heredity and
reproduction.
Increasing plant and animal food production, diagnosing and treating diseases
through medical advancement, and producing useful drugs and vaccines for the
Sars-Cov2, are the major focus on the importance of genetic engineering.

THE DIFFERENT TOOLS USED IN GENETIC ENGINEERING

Here are tools that are commonly used in genetic engineering:
● Polymerase Chain Reaction (PCR) is efficient technique because it multiplies
the DNA exponentially for 25 to 75 cycles. A cycle takes only a minute, and each
new segment of DNA that is made can serve as a template for new ones. This
technique is used in molecular biology to amplify a single copy or a few copies of
a segment of DNA across several orders of magnitude, generating thousands to
millions of copies of a particular DNA sequence. Developed in 1983 by Kary
Mullis, PCR is now a common technique used in clinical and research
laboratories for various applications (Javed, 2017).

Figure 2: Polymerase Chain Reaction

Image Source: https://explorebiotech.com/10-tools-for-genetic-engineering/

● Restriction Enzymes (Molecular Scissor) are enzymes that create one incision
on each of the two strands of DNA at specific locations based on the nucleotide
sequence. DNA cut with a restriction enzyme produces many smaller fragments
of varying sizes. These can be separated using gel electrophoresis or
chromatography. Furthermore, hundreds of different restriction enzymes
isolated from many different bacteria strains are capable of cutting DNA at a
distinct site. Restriction Enzymes was isolated in 1970 by Hindll. He also did the
subsequent discovery and characterization of numerous restriction
endonucleases (Hitendra, 2018).
 Figure 2: Restriction Enzymes
Image Source: https://explorebiotech.com/10-tools-for-genetic-engineering/

● Gel Electrophoresis is used for various purposes, from viewing cut DNA to
detecting DNA inserts and knockouts. It is also used to estimate the molecular
weight of protein and nucleic acids, purification of isolated proteins, monitoring
changes of protein content in body fluids, blotting application, and many more.
Purifying DNA from cell culture or cutting it using restriction enzymes would not
be of much use if we could not visualize the DNA that is, find a way to view
whether or not your extract contains anything or what size fragments you have
cut it into. One way to do this is by gel electrophoresis (Tapeshwar, 2015).

Figure 3: Gel Electrophoresis

Image Source: https://explorebiotech.com/10-tools-for-genetic-engineering/
● DNA Ligase are enzymes that can create covalent bonds between nucleotide
chains. The enzymes DNA polymerase I and polynucleotide kinase are also
crucial in filling in gaps, or phosphorylating the 5′ ends, respectively. The DNA
ligase used in molecular cloning differs in their abilities to ligate noncanonical
substrates, such as blunt-ended duplexes DNA: RNA hybrid or ss DNAs. In
genetic research, DNA Ligase are often utilized to link two or more individual
strands of DNA, to create a recombinant strand, or close a circular strand that
has been cut with restriction enzymes (Faraza ,2017).

Figure 4: DNA Ligation Process

Image Source: https://explorebiotech.com/10-tools-for-genetic-engineering/

● According to Faraza (2017), polymerases are collectively referred as the groups

of enzymes that catalyze nucleic acid molecules’ synthesis. It is customary to use
the name of the nucleic acid template on which the polymerase acts. The three
critical polymerases are given below.
 Figure 5: Polymerizing and Proofreading
Image Source: https://explorebiotech.com/10-tools-for-genetic-engineering/

o DNA-dependent DNA polymerase that replicates DNA from DNA.

o RNA-dependent DNA polymerase (reverse transcriptase) that transcribes
DNA from RNA. o DNA-dependent RNA polymerase that transcribes RNA
from DNA

Figure 6: Plasmid Map

Image Source: https://explorebiotech.com/10-tools-for-genetic-engineering/

● Prokaryotic Host are able to multiply their plasmids (along with foreign DNA)
also multiply to produce millions of copies, referred to as a colony or in a short
clone. The term ‘clone’ broadly refers to a mass of cells, organisms, or genes that
 results from the multiplication of a single cell, organism, or gene. Furthermore,
the bacteria Escherichia coli, was the first organism used in the DNA technology
experiments and continues to be the host of choice by many workers.
Undoubtedly, E.coli, the simplest Gram-negative bacterium (a common
bacterium of human and animal intestines), has played a key role in the
development of present-day biotechnology. Under a suitable environment, the
number of E. coli can double every 20 minutes (Faraza ,2017).

• Eukaryotic Host are preferred to produce human proteins since these hosts with
complex structures (with distinct organelles) are more suitable to synthesize
complex proteins. The most commonly used eukaryotic organism is the yeast,
Saccharomyces cerevisiae. It is a non-pathogenic organism routinely used in the
brewing and baking industry. Certain fungi have also been used in gene cloning
experiments (Faraza ,2017).

● Selection of Small Self-Replicating DNA

Small circular pieces of DNA that are not part of a bacterial genome, but are
capable of self-replication, are known as plasmids. Plasmids are often used as
vectors to transport genes between microorganisms. In biotechnology, once the
gene of interest has been amplified and restriction enzymes cut both the gene
and plasmid, they are ligated together, generating what is known as recombinant
DNA. Viral (bacteriophage) DNA can also be used as a vector, as can cosmids,
recombinant plasmids containing bacteriophage genes (Faraza ,2017).
 Figure 7: Plasmid Map
Image Source: https://explorebiotech.com/10-tools-for-genetic-engineering/

● Methods to Select Transgenic Organisms

According to Faraza (2017), not all cells will take up DNA during transformation.
Therefore, it is essential to identify the cells that transform and those that have
not. Generally, plasmids carry genes for antibiotic resistance, and transgenic
cells can be selected based on the expression of those genes and their ability to
grow on media containing that antibiotic. Alternative selection methods depend
on the presence of other reporter proteins such as the x-gal/lacZ system or green
fluorescence protein, which allow selection based on color and fluorescence,
respectively.
 Figure 8: Selection of Transgenic Organisms
Image Source: https://explorebiotech.com/10-tools-for-genetic-engineering/