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Biotechnology
Quarter 3 – Module 3:
Steps in Recombinant DNA
Technology

DIVISION OF ANGELES CITY

Biotechnology– Grade 8
Alternative Delivery Mode
Quarter 3 – Module 3: Steps in Recombinant DNA Technology
First Edition, 2021

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Published by the Department of Education

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Assistant Regional Director: Rhoda T. Razon EdD, CESO V

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Editors: Sherilyne L. Reyes, Jennifer Praza, Edgardo D. Cortez,
Jenny S. Tongol, Edythe Hipolito
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Management Team: May B. Eclar PhD, CESO V
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Rochella C. David
Emily F. Sarmiento PhD
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Biotechnology
Quarter 3 – Module 3:
Steps in Recombinant DNA
Technology
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 self-check
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

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 This module was designed and written with you in mind. It is here to help you
master the steps in Recombinant DNA Technology. 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 about:

● Steps in Recombinant DNA Technology

After going through this module, you are expected to:

1. Outline the steps in Recombinant DNA Technology

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 What I Know

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

best answer.

1. Which refers to the combination of two DNA strands that are constructed
artificially?
a. Genetic Material
b. Recombinant Cells
c. Recombinant DNA
d. Restriction Enzymes

2. Which among the following is the Blueprint of Life?

a. Cell
b. DNA
c. Protein
d. RNA

3. Which refers to the small accessory ring of the DNA in some bacteria? a.
Interferon
b. Plasmid
c. Restriction enzymes
d. Vector

4. What molecule is being used to cut a specific area of the DNA?

a. Agarose Gel
b. Plasmid
c. Recombinant Cells
d. Restriction enzymes

5. An organism that contains genetic material from two different organisms is

called ___________. a. Clone
b. GMO
c. Mutant
d. Restriction enzymes

6. What technique is being used to make a million copies of a sample DNA?

a. DNA Fingerprinting
b. Gel Electrophoresis
c. Gene Therapy
d. Polymerase Chain Reaction

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7. Which among the following does NOT describe the principle of Gel
Electrophoresis?
a. DNA fragments are separated on the basis of size.
b. The DNA fragments will move towards the negative charge.
c. The smallest DNA fragments will move faster than the larger DNA
fragments.
d. DNA fragments are injected into wells and an electric current is applied
along with the gel.

8. What equipment is used to amplify the DNA?

a. Centrifuge Machine
b. Microscope
c. Pipette
d. Thermal Cycler

9. Which among the following serves as a starting point for DNA synthesis?
a. DNA polymerase
b. Primer
c. Restriction enzymes
d. Taq polymerase

10. What organism is being used to transfer foreign genetic material into a cell?
a. DNA
b. Plasmid
c. Restriction enzymes
d. Vector

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 Lesson
Steps in Recombinant DNA
1 Technology
What’s In

Activity 1
Direction: Write True if the statement is correct and False if incorrect.

1. Genetic Engineering is the artificial manipulation of DNA or other nucleic acid

molecules in order to modify an organism or population of organisms.
2. Restriction enzyme was discovered by Werner Arber.
3. Genes are small rings of DNA.
4. DNA technology makes it possible to clone genes for basic research and
commercial applications.
5. Restriction enzymes cut the DNA into a particular site.
In our previous lesson, we learned how genetic materials are manipulated.
One of the ways is through Genetic engineering. In Genetic engineering, genes are
manipulated for practical purposes.

Genetic Engineering

DNA technology has launched a revolution in Biotechnology. DNA technology (via

gene manipulation) makes it possible to clone genes for basic research and
commercial applications. DNA technology is applied to areas ranging from
agriculture to criminal law. One example of DNA technology is Genetic engineering.
Genetic Engineering is the the artificial manipulation, modification, and
recombination of DNA or other nucleic acid molecules in order to modify an organism
or population of organisms. In the latter part of the 20th century, however, the term
came to refer more specifically to methods of recombinant DNA technology, in which
DNA molecules from two or more sources are combined either within cells or in vitro
and are then inserted into host organisms in which they are able to propagate.

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 The possibility for recombinant DNA technology emerged with the discovery of
restriction enzymes in 1968 by Swiss microbiologist Werner Arber.

Most recombinant DNA technology involves the insertion of foreign genes into the
plasmids of common laboratory strains of bacteria. Plasmids are small rings of DNA;
they are not part of the bacterium’s chromosome. Nonetheless, they are capable of
directing protein synthesis, and, like chromosomal DNA, they are reproduced and
passed on to the bacterium’s progeny. Thus, by incorporating foreign DNA (for
example, a mammalian gene) into a bacterium, researchers can obtain an almost
limitless number of copies of the inserted gene. Furthermore, if the inserted gene is
operative (if it directs protein synthesis), the modified bacteria will produce the
protein specified by the foreign DNA. Editors of Encyclopedia Britannica (2020).

What’s New
Activity 1. Steps in Recombinant DNA Technology
Directions: Arrange the steps in Recombinant DNA Technology in chronological
order. Use numbers 1-7.
A. Amplification Using PCR
B. Isolation of Recombinant Cell
C. Isolation of Genetic Material
D. Obtaining or culturing the Foreign Gene product.
E. Ligation of DNA Molecules.
F. Restriction Enzymes Digestion
G. Insertion of Recombinant DNA into Host

What is It

In this lesson, we shall outline the main steps in Recombinant

DNA Technology and the importance of Recombinant DNA Technology .

Recombinant DNA Technology

Recombinant DNA technology refers to the joining together of DNA molecules

from two different species that are inserted into a host organism to produce new
genetic combinations that are of value to science, medicine, agriculture, and
industry. Recombinant DNA (rDNA), on the other hand, is the general name for a

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piece of DNA that has been created by the combination of at least two different DNA
strands. They are DNA molecules formed by laboratory methods of genetic
recombination to bring together genetic material from multiple sources, creating
sequences that would not otherwise be found in the genome. Aryal (2018).

Steps of Genetic Recombination Technology

1. Isolation of Genetic Material - Since DNA exists within the cell membrane
along with other macromolecules such as RNA, polysaccharides, proteins, and
lipids, it must be separated and purified which involves enzymes such as
restriction enzymes.

2. Restriction Enzymes Digestion - The technique ‘Agarose Gel Electrophoresis’

reveals the progress of the restriction enzyme digestion. This technique involves
running out the DNA on an agarose gel. On the application of current, the
negatively charged DNA travels to the positive electrode and is separated out
based on size. This allows separating and cutting out the digested DNA
fragments. The vector DNA is also processed using the same procedure.

3. Amplification Using PCR - Polymerase Chain Reaction or PCR is a method of

making multiple copies of a DNA sequence using the enzyme – DNA polymerase
in vitro. It helps to amplify a single copy or a few copies of DNA into thousands
to millions of copies. PCR reactions are run on thermal cyclers using the
following components: 1.)Template – DNA to be amplified 2.)Primers – small,
chemically synthesized oligonucleotides that are complementary to a region of
the DNA. 3.) Enzyme – DNA polymerase 4.)Nucleotides – needed to extend the
primers by the enzyme. 5.) The cut fragments of DNA can be amplified using PCR
and then ligated with the cut vector.

4. Ligation of DNA Molecules – The purified DNA and the vector of interest are cut
with the same restriction enzyme. This gives us the cut fragment of DNA and the
cut vector that is now open. The process of joining these two pieces together
using the enzyme DNA ligase is ligation. The resulting DNA molecule is a hybrid
of two DNA molecules – the interest molecule and the vector. In the terminology
of genetics this intermixing of different DNA strands is called recombination.
Hence, this new hybrid DNA molecule is also called a recombinant DNA molecule
and the technology is referred to as the recombinant DNA technology.

5. Insertion of Recombinant DNA into Host - In this step, the recombinant DNA
is introduced into a recipient host cell mostly, a bacterial cell. This process is
called transformation. Bacterial cells do not accept foreign DNA easily. Therefore,
they are treated to make them competent to accept new DNA. The processes used
may be thermal shock, Ca++ ion treatment, and electroporation.

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6. Isolation of Recombinant Cells-The transformation process generates a mixed
population of transformed and non-transformed host cells. The selection process
involves filtering the transformed host cells only. For isolation of recombinant
cells from non-recombinant cells, a marker gene of the plasmid vector is
employed.
7. Obtaining or culturing the Foreign Gene product - When you insert a piece of
alien DNA into a cloning vector and transfer it into a bacterial cell, the alien DNA
gets multiplied. The ultimate aim is to produce a desirable protein expression.
The cells harboring cloned genes of interest are grown on a small scale in the
laboratory. These cell cultures are used for extracting the desired protein using
various separation techniques.

Recombinant Human Growth Hormone

Source: https://simplebiologyy.blogspot.com/2016/02/process-of-
recombinantdna-technology-genetic-engineering.html

Importance of Recombinant DNA Technology

Recombinant DNA technology is playing a vital role in improving health

conditions by developing new vaccines and pharmaceuticals. The treatment
strategies are also improved by developing diagnostic kits, monitoring devices, and
new therapeutic approaches. Synthesis of synthetic human insulin and
erythropoietin by genetically modified bacteria and the production of new types of
experimental mutant mice for research purposes are some one of the leading
examples of genetic engineering in health. Likewise, genetic engineering strategies

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have been employed to tackle environmental issues such as converting wastes into
biofuels and bioethanol, cleaning the oil spills, carbon, and other toxic wastes, and
detecting arsenic and other contaminants in drinking water. The genetically modified
microbes are also effectively used in biomining and bioremediation.

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