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Biotech8 q3 Mod3 StepsinRecombinantDNATechnology v3
Biotech8 q3 Mod3 StepsinRecombinantDNATechnology v3
Biotechnology
Quarter 3 – Module 3:
Steps in Recombinant DNA
Technology
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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-by-
step as you discover and understand the lesson prepared for you.
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.
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What I Need to Know
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.
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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
3. Which refers to the small accessory ring of the DNA in some bacteria?
a. Interferon
b. Plasmid
c. Restriction enzymes
d. Vector
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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.
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.
Genetic Engineering
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
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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
What is It
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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.
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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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.
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detecting arsenic and other contaminants in drinking water. The genetically modified
microbes are also effectively used in biomining and bioremediation.
What’s More
Guide Questions
1. What is the first step in Recombinant DNA Technology?
2. In what step does the cut fragment of DNA and the cut vector are joined
together?
3. What is the final step in Recombinant DNA Technology?
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Activity 2. Modeling Bacteria Transformation
Directions: Using the word choices provided in the boxes, fill in the numbered boxes
with the steps of bacteria transformation and the lettered lines with the name of the
structure next to them.
Source:https://www.teachengineering.org/activities/view/uoh_genetic_lesson01_a
ctivity1.
Guide Questions
1. What is the role of restriction enzymes in Recombinant DNA Technology?
2. What is the function of the DNA Ligase?
3. What is a Plasmid?
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Activity 3 . Recombinant DNA Technology
Directions: Read the choices from each numbered item in Column A and identify
which is NOT included from these groups. Then classify it by choosing the correct
answer in Column B.
Note: To get one (1) point from this activity, two (2) responses must be answered
correctly.
Column A Column B
1. a. Primers A. Isolation of Genetic Material
b. DNA Polymerase
c. Agarose Gel
d. PCR B. Restriction Enzymes Digestion
2. a. Transformation
b. Ligation C. Amplification Using PCR
c. Recombinant DNA
d. Ligase
D. Ligation of DNA Molecules
3. a. Gel Electrophoresis
b. Protein expression
c. Positive electrode
d. Agarose Gel E. Insertion of Recombinant DNA
into Host
4. a. Marker gene is employed.
b. Filtering of the transformed host cell.
c. Negatively charged DNA travels to the F. Isolation of Recombinant Cells
positive electrode.
d. Mixed population of transformed and
non-transformed cells.
G. Obtaining or culturing the
5. a. DNA is separated based on size. Foreign Gene product
b. Cutting out of digested DNA
fragments.
c. Negatively charged DNA travels to the
positive electrode.
d. Amplify a single copy of DNA into
thousands or millions
Guide Questions
1. What are Primers?
2. What is the function of the PCR in Recombinant DNA Technology?
3. What is the charge of the DNA?
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Activity 4. Complete the Steps in Recombinant DNA Technology
Directions: Complete the figure below by supplying the missing Step in
Recombinant DNA Technology.
2.
4.
5.
7.
Guide Questions
1. What enzyme is used in DNA Ligation?
2. In what step in Recombinant DNA does Transformation occur?
3. What is the function of the Gel Electrophoresis in Recombinant DNA
Technology?
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Activity 5. Describing the steps in Recombinant DNA Technology
Directions: Identify which step in Recombinant DNA Technology is involved.
Guide Questions
1. What equipment makes multiple copies of the DNA?
2. How do DNA fragments separate in Gel Electrophoresis?
3. What enzyme is being used to separate and purify the DNA from the cell?
Eli Lilly began producing insulin from animal pancreas but fell short of the
demand, and the potency varied up to 25% per lot. The development of an isoelectric
precipitation method led to purer and more potent animal insulin, decreasing the
variation between lots to 10%. These discoveries led to the introduction of longer-
acting animal insulins in the market. Protamine zinc insulin lasted 24–36 hours.
Isophane neutral protamine Hagedorn lasted 24 hours and could be mixed with
regular insulin. The pharmacokinetics and effects of amorphous lente insulin
(semilente, lente, and ultralente) depended on the proportion of zinc. In 1978, the
first recombinant DNA human insulin was prepared by David Goeddel and his
colleagues (of Genentech) by utilizing and combining the insulin A- and B- chains
expressed in Escherichia coli. Thereafter, Genentech and Lilly signed an agreement
to commercialize rDNA insulin. In 1982, the first insulin utilizing rDNA technology,
Humulin® R (rapid) and N (NPH, intermediate-acting), were marketed.
Guide Questions
1. Are you in favor of using the animal pancreas to replace the insulin in the
human body? Why?
2. Based on the article, how does Recombinant DNA benefit humans?
3. What is Recombinant DNA Technology?
4. What is the function of a vector?
5. What are the common vectors in Recombinant DNA Technology?
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What I Have Learned
1. The general name for a piece of DNA that has been created by the combination
of at least two strands of DNA is called _____________.
2. The process of introducing recombinant DNA into a recipient host cell is called
_____________ .
3. Agarose Gel Electrophoresis involves running out the DNA on an
_____________.
4. The process of joining the cut fragment of DNA and the cut vector together
using an enzyme is called_____________ .
5. The _____________ is a method of making multiple copies of a DNA sequence
using an enzyme.
6. Recombinant DNA technology refers to the joining together of _____________
from two different _____________ that are inserted into a host organism to
produce new genetic combinations.
7. Ligation of DNA molecules uses_____________ to cut the vector.
8. An enzyme called _____________ helps to amplify a single copy or a few copies
of DNA into thousands to millions of copies.
9. PCR reactions are run on _____________ to amplify a single copy or a few copies
of DNA.
10. The small circular molecules which act as carriers for the DNA fragments are
called _____________.
What I Can Do
1. Health
2. Food
3. Environment
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Assessment
Directions: Read each statement carefully. Choose the letter of the correct
answer.
1. Which enzyme is being used to amplify the DNA?
a. Endonuclease
b. DNA Helicase
c. DNA Ligase
d. DNA Polymerase
For question nos. 3-9, Arrange in order the steps in Recombinant DNA Technology.
Use letters a to g
3. Amplification Using PCR
4. Isolation of Genetic Material
5. Insertion of Recombinant DNA into Host
6. Obtaining or culturing the Foreign Gene product.
7. Ligation of DNA Molecules.
8. Restriction Enzymes Digestion
9. Isolation of Recombinant Cells
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For question nos. 12-15. Identify the steps in Recombinant DNA Technology that are
being described in each statement. The choices are as follows:
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Additional Activity
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What I Know What’s In What’s New
1. C 6. D 1. True Activity 1
2.B 7. B 2.True 1. Isolation of Genetic Material
3.B 8. D 3.False 2. Restriction Enzymes Digestion
4.True 3. Amplification Using PCR
4.D 9. B
5.True 4. Ligation of DNA Molecules.
5.B 10. D
5. Insertion of Recombinant DNA into Host.
6. Isolation of Recombinant Cells
7. Obtaining or culturing the Foreign Gene
product.
What’s More What’s More
Activity 1 Activity 4
DNA Ligase - joins two pieces of DNA. 2. Restriction Enzymes Digestion
DNA Polymerase - makes multiple copies of 4. Ligation of DNA Molecules.
a DNA 5. Insertion of Recombinant DNA into Host .
Similarity: Catalyst for Recombinant DNA 7. Obtaining or culturing the Foreign Gene
Technology product.
Guide Quest ions Guide Quest ions
1. Isolation of Genetic Material 1. DNA Ligase
2. Ligation of DNA Molecules. 2. Insertion of Recombinant DNA into Host
3. Obtaining or culturing the Foreign Gene Cell
product. 3. Gel Electrophoresis runs out the DNA on
Activity 2 an agarose gel.
A. Plasmid
Activity 5
B. Foreign DNA with desired genes
1. Restriction Enzymes Digestion
C. Recombinant DNA
2. Obtaining or culturing the Foreign Gene
1. Plasmid cut with restriction Enzymes product.
2. DNA Ligase joins sticky ends 3. Isolation of Genetic Material
3. Bacteria transformed 4. Insertion of Recombinant DNA into Host .
Guide Quest ions 5. Amplification Using PCR
1.Restriction Enzymes separate the DNA Guide Quest ions
from the cell. 1. Thermal Cycler
2.DNA Ligase joins two pieces of DNA. 2. Based on size
3.Plasmid is the accessory ring of a bacterial 3. Restriction Enzymes
chromosome.
Activity 6
Activity 3
Answers may vary
1. C, C
Possible answer :
2. A, D
1. Yes, it helps to solve the problem on
3. B, B
insufficient supply of insulin.
4. C, F
2. Recombinant DNA can produce artificial
5. D, B
human insulin.
Guide Quest ions
3.Recombinant DNA technology is the
1. Primers are small, chemically
joining together of DNA molecules from two
synthesized oligonucleotides.
different species that are inserted into a host
2. PCR makes a multiple copies of a DNA
organism to produce new genetic
sequence.
3. Negatively charged. combinations.
4. Vector is use to transfer foreign genetic
material into a cell.
5. Bacteria and Virus
Answer Key
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What I Have Learned What Can I Do Assessment
1. Recombinant Answers may vary 1.D 11. A
DNA Possible answer: 2.C 12. G
2. Transformation Health- Artificial human insulin 3.C 13. D
3. Agarose gel helps Diabetic people. 4.A 14. B
4. Ligation Food- Golden Rice with a 5.E 15. C
5. PCR enhanced nutritional value. 6.G
6. DNA molecules,
Environment- Pest resistant crops 7.D
species
uses less pesticide, therefore less 8.B
7. DNA ligase
harm to the environment 9.F
8. DNA polymerase
9. Thermal Cycler 10.B
10. Vector
Additional Activity
Answers may vary
Possible answer:
Introduction of genetically modified organisms which are product of Recombinant DNA
Technology might change the gene frequency of a population that could lead to extinction of
some organisms.
References
Aryal, Sagar, and Rashid Eltayeb. 2020. “Recombinant DNA Technology- Steps,
Applications and Limitations: Molecular Biology.” Microbe Notes.
https://microbenotes.com/recombinant-dna-technology-steps-applications-
and-limitations/.
Griffiths AJF, Miller JH, Suzuki DT, et al. 2020. An Introduction to Genetic Analysis.
7th edition. New York: W. H. Freeman; 2000.
https://www.ncbi.nlm.nih.gov/books/NBK21881/
Quianzon, Celeste C., and Issam Cheikh. 2012. "History of insulin." Journal of
community hospital internal medicine perspectives 2, no. 2: 18701.
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