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GENERAL BIOLOGY 2
Third Quarter
Module No. 1 of 3
GENETIC ENGINEERING
Writer: Roland R. Agra

HONOR CODE
AS A MEMBER OF THE NAMUAC ACADEMY EAGLES FAMILY, I WILL CONDUCT
MYSELF WITH INTEGRITY & SINCERITY AT ALL TIMES, DEMONSTRATE COMPASSION &
JUSTICE IN ALL MY ACTIONS, UPHOLD THE VALUE OF EXCELLENCE, AND ABIDE BY THE
EXPECTATIONS SET FORTH IN THE STUDENT HANDBOOK.
I MAKE THIS PLEDGE IN THE SPIRIT OF HONOR & TRUST.

PERFORMANCE TASK IN GENERAL BIOLOGY 2

PERFORMANCE Make a research paper/case study/poster on genetic diseases
STANDARDS
GOAL To raise awareness on genetic disorders
ROLE Visual artist/designer, nurse, health advocate/educator
AUDIENCE The general public
SITUATION Research shows that there is increase in the occurrence of genetic
disorders among newly born infants whose mothers did not undergo
regular monthly check-ups or newborn screening. You were tasked by
the Department of Health to conduct an information drive what are the
common genetic diseases and how can they be managed or prevented.
PRODUCT Multimedia presentation
STANDARDS Accuracy, organization of ideas, clarity, use of appropriate illustration

21ST CENTURY SKILLS CORE VALUE TASK

CRITICAL THINKING Excellence How to create a multimedia presentation based on
the standards
CREATIVITY Excellence To present a well-thought MMP
COLLABORATION Justice The students will work together as members of the
team to prepare the MMP.
CROSS CULTURAL
UNDERSTANDING
COMPUTER/ICT Excellence Using ICT in looking for information about genetic
disorders
CAREER/SELF RELIANCE
COMMUNICATION Compassion Communicating the MMP to the intended audience

SCORING RUBRIC FOR THE PERFORMANCE TASK

CRITERIA 4 3 2 1
Accuracy All concepts were All concepts were All concepts were All concepts were
accurate and accurate and accurate and accurate and
showed deep showed considerable showed showed limited
understanding of understanding of the considerable understanding of
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the topic. Causes topic. Causes of understanding of the topic. Causes

of genetic genetic disorders the topic. Causes of genetic
disorders were were extensively of genetic disorders were
extensively discussed and disorders were discussed and
discussed and management discussed and management
management measures were management measures were
measures were presented measures were presented.
presented accurately. presented.
accurately.
Organization of Concepts were Concepts were Concepts were Concepts were
Ideas relevant, accurate relevant and relevant and relevant but not
and effective in accurate but the somewhat accurate. The
conveying the information was not accurate. The information was
information. conveyed effectively. information was not conveyed
not conveyed effectively.
effectively.
Clarity Established a clear Established a clear Established a clear Failed to
purpose that is purpose that is quite purpose that is establish a clear
relevant to the goal relevant to the goal quite relevant to purpose relevant
and demonstrated and demonstrated a the goal but did to the goal but did
a clear clear understanding not demonstrate a not demonstrate
understanding of of the topic. clear a clear
the topic. understanding of understanding of
the topic. the topic.
Use of The illustrations The illustrations The illustrations The illustrations
appropriate used were used were used were used were
illustrations appropriate and appropriate and appropriate and somewhat
visually appealing somewhat visually not visually appropriate and
and to the topics appealing and to appealing and to not visually
discussed. the topics discussed. the topics appealing and to
discussed. the topics
discussed.

EXPECTATION
S
After going through this module, you are expected to:
1. Outline the processes involved in genetic engineering. (STEM_BIO11/12-IIIa-b-6)
2. Discuss the applications of recombinant DNA. (STEM_BIO11/12-IIIa-b-7)

PRE-TEST

MULTIPLE CHOICE. Read and understand each item and choose the letter of the correct answer.
Write your answers on a separate sheet of paper.

1. What is the process of making changes on the genetic code of an organism?

a. Civil engineering c. Chemical engineering
b. Genetic engineering d. Biological engineering

2. What is the process of modifying the genes of organisms for practical purposes?

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a. RNA splicing b. Polyadenylation c. DNA transcription d. DNA

recombination

3. What organisms that are normally harmless but may help spread infection by transferring the
genetic material from one host to another?
a. Vectors b. Plasmid c. Allergens d. Transgenic plants

4. What process in which a selected portion of the foreign DNA is inserted into a small circular
DNA molecule found in bacteria?
a. Transduction b. Transformation c. Electroporation d. Vectorless gene transfer

5. What process wherein genetically engineered bacteriophages are introduced into the cell to
create the desired recombinant DNA?
a. Transduction b. Transformation c. Electroporation d. Vectorless gene transfer

6. At what level are gene products manipulated in genetic engineering?

a. Amino acid b. DNA c. Protein d. RNA

7. An organism that contains genes from other organisms is called________.

a. Macrotrans b. recomorgi c. transgenic d. transplant

8. Which microorganisms used in the application of recombinant DNA?

a. Algae b. Bacteria c. Fungi d. Virus

9. Which is a product of genetically modified organism or GMO in plants?

a. BT corn b. Cheese c. Gene therapy d. Vaccines

10. What is the application of recombinant DNA in the field of agriculture?

a. Maintain good health c. Improved nutritional value
b. To treat human disease d. Developed insect-resistant plant

OVERVIEW

Genetic engineering is a process of making changes on the genetic code of an organism. Its
goal is to add one or more new traits that are not normally found in that organism. Through advanced
studies in the structure of DNA and its chemical properties, scientists have been able to employ
different techniques to extract, cut, and make unlimited copies of DNA.

DNA recombination is a process of modifying the genes of organisms for practical purposes. It
is done when a piece of DNA is combined with another DNA from another source. The resulting genetic
product is called recombinant DNA. With this process, organisms get to have traits that are not normally
found in their species.

Several scientific advancements have led to many genetic engineering techniques that are very
beneficial to us. It is now possible to transfer DNA sequences from one organism to another. American
researcher Steven Howell and his associates at the University of California in San Diego learned that
even genes from two or more different organisms can be made to work together. Howell’s team tried to
isolate the gene for luciferase-an enzyme that allows fireflies to glow-and insert it into tobacco cells.

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When the gene was activated from the recombinant cells, the plants glowed in the dark. This
means that the basic mechanisms of gene expression are shared by both plants and animals.

LESSON PROPER

GENETICS. It is a field of biology that

studies how traits are passed from parents to their
offspring. It is the study of how living things receive
common traits from previous generations. These
traits are described by the genetic information
carried by a molecule called DNA. The instructions
for constructing and operating an organism are
contained in the organism’s DNA.

DNA. Every living organism on earth has

DNA in its cells. Deoxyribonucleic acid (DNA) is a
molecule that contains the biological instructions that make each species unique. DNA, along with the
instructions it contains, is passed from adult organisms to their offspring during reproduction.

GENETIC ENGINEERING. It is
artificially copying a piece of DNA from
one organism and joining this copy of DNA
into the DNA of another organism. It is
defined as the direct manipulation of an
organism’s genes including heritable and
nonheritable recombinant DNA constructs.
Genetic engineering involves the use of
molecular techniques to modify the traits
of a target organism.

The modification of traits may

involve introduction of new traits into an
organism; enhancement of a present trait
by increasing the expression of the desired gene; or enhancement of a present trait by disrupting the
inhibition of the desired genes’ expression.

RECOMBINANT DNA TECHNOLOGY. It is one technique wherein a gene of interest from one
organism is inserted into the genome of another. This involves gene cloning using a bacterial plasmid
as a vector. It is widely used in improving crop varieties.

Recombinant DNA technologies, developed in the latter half of the twentieth century, include
the chemical splicing (recombination) of different strands of DNA generally using either bacteria (such
as Escherichia coli) or bacteriophages (viruses that infect bacteria), or by direct microinjection.

GENERAL OUTLINE ON THE FORMATION OF RECOMBINANT DNA

1. Isolation of a specific gene from donor. The first step involves breaking open the cells of
the donor to release the DNA and isolate the gene of interest.

a. Cells are broken open using chemicals and enzymes. Donor DNA is extracted.
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b. Genetic probe is added. A DNA probe consists of a small fragment of DNA labelled with
an enzyme, a radioactive tag or a fluorescent dye tag. The probe will bind to a
complementary DNA sequence by base pairing.
c. Reveal position of the gene of interest.
2.

Cutting or cleavage of DNA by restriction enzymes (REs)

 The DNA from the bacterial cell is released.
 A bacterial cell contains a
circular loop of DNA called
a plasmid.
 The plasmid is isolated
from the bacterial cell.
 The plasmid will act as a
vector for carrying a new
gene i.e. the gene from the
donor will be inserted into
the plasmid DNA.
 The donor DNA and
plasmid DNA are cut using
enzymes called restriction
enzymes which recognize specific base sequences and cats as a molecular scissors to cut
DNA strand within the recognition sequence.
 The donor DNA and plasmid DNA are cut using the same restriction enzymes.
 Restriction enzymes cut the DNA from the donor and the plasmid at specific points.
 The cut ends have sticky ends which have unpaired bases

3. Ligation is re-joining cut fragments of DNA and forming artificial recombinant molecules. It is
joining together of the gene of interest (eg. from animal) with the vector (cut bacterial plasmid)

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4. Transformation- the transfer of the recombinant plasmid into a host cell that would carry out
replication to make huge copies of the recombined plasmid recombinant DNA introduced into
bacterial cell.

Ways in which Plasmids may be introduced into Host Organisms

a. Biolistics. In this technique, a “gene gun” is used to fire DNA-coated pellets on plant tissues. Cells
that survive the bombardment and are able to take up the expression plasmid coated pellets and
acquire the ability to express the designed protein.

b. Plasmid insertion by Heat Shock Treatment. It is a process used to transfer plasmid DNA into
bacteria. The target cells are pre-treated before the procedure to increase the pore sizes of their
plasma membranes. This pretreatment (usually with CaCl 2) is said to make the cells “competent”
for accepting the plasmid DNA. After the cells are made competent, they are incubated with the
desired plasmid at about 4°C for about 30 minutes. The plasmids concentrate near the cells during
this time. Afterwards, 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 is believed to
increase and decrease 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”.

c. 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 insertion of genes into mammalian cells.

Some Methods to Screen Recombinant Cells

a. 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 (e.g., AMP
ampicillin resistance gene) is included in the plasmid DNA. 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.

b. Selection of transformed cells with the desired gene. Certain inserted genes within the plasmids
provide visible proof of their presence. These include the antibiotic resistant genes that allow for the
selection of the transformed cells within the solution. Some inserted genes also produce colored
(e.g., chromogenic proteins) or fluorescent products (e.g., GFP) that label the colonies/cells with the
inserted gene. In some cases, the location of the cloning site within the plasmid is in the middle of a
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gene (i.e., β-galactosidase, lacZ) that generates a (blue) colored product in the presence of a
substrate (i.e., isopropyl β-D-1 thiogalactopyranoside, or IPTG). Cells transformed with these
“empty” plasmids will turn blue in the presence of IPTG. Insertion of a gene in the cloning site
disrupts the sequence of the β-galactosidase gene and prevents the generation of the colored
product in the presence of the substrate. Cells transformed with the disrupted β-galactosidase gene
will remain “white” in the presence of IPTG. This “blue-white screening” protocol is thus able to
screen for cells that were transformed with the desired gene in the cloning site.

c. Polymerase chain reaction detection of plasmid DNA Alternatively, the presence of the desired gene
in the inserted plasmids may be confirmed using PCR amplification. PCR reactions specific for the
desired gene may be done using DNA from cells. Amplification of the expected product would
confirm the presence of the gene within the samples. PCR reactions specific for plasmid sequences
will also confirm/identify the type of plasmid used for the transformation.

5. Expression. Plasmid will produce the

polypeptide coded for by the donor DNA.
 Bacterial cell reproduces by Binary
Fission
 Bacterial cell produces the
polypeptide coded for by the donor
DNA

TRANSGENIC ORGANISM. These are

organisms altered by genetic engineering.
The genetic material can be changed by
other than random natural breeding or by
gene transfer which is the moving of a gene
from one organism to another.

GENETICALLY MODIFIED ORGANISM.

Genetically modified organisms (GMOs) are
living organisms whose genetic material has been artificially manipulated in a laboratory through
genetic engineering. This creates combinations of plant, animal, bacteria, and virus genes that do
not occur in nature or through traditional crossbreeding methods.

RECOMBINANT DNA. The altered DNA is

called recombinant DNA in which
molecules of DNA 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 technology has made it possible to
isolate one gene or any other segment of
DNA, enabling researchers to determine
its nucleotide sequence, study its
transcripts, mutate it in highly specific ways, and reinsert the modified sequence into a living
organism.
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Applications of Recombinant DNA

1. Medicine. Genetic engineering has been used to mass-produce insulin, human growth
hormones, follistim (for treating infertility), human albumin, monoclonal antibodies, antihemophilic
factors, vaccines, and many other drugs.

2. Industrial Applications.
Genetically designed
bacteria are put into use for
generating industrial
chemicals. A variety of
organic chemicals can be
synthesized at large scale
with the help of genetically engineered microorganisms. Glucose can be synthesized from
sucrose with the help of enzymes obtained from genetically modified organisms.
a. Bacteria that metabolize petroleum and other toxic materials have been developed.
b. Scorpion toxin gene has been inserted into autographa California multicapsid nuclear
polyhedrosis virus (AcMNPV) which kills cabbage looper and reduce crop damages.
c. Development of new strains for additional bioprocesses
3. Agricultural Applications. An important application of recombinant DNA technology is to alter the
genotype of crop plants to make them more productive, nutritious, rich in proteins, disease
resistant, and less fertilizer consuming. Recombinant DNA technology and tissue culture
techniques can produce high yielding cereals, pulses and vegetable crops.
Some plants have been genetically programmed to yield high protein grains that could
show resistance to heat, moisture and diseases.
Scientists have developed transgenic potato, tobacco, cotton, corn, strawberry, rape seeds
that are resistant to insect pests and certain weedicides.
a. Genes of interest is inserted into plant with Ti plasmid obtained from Agrobacterium
tumefaciens.
b. Pseudomonas syringae that protects plants from frost damage is used against plant
frost damage because they lack the protein that induce the formation of ice-crystals.
c. Insertion of Bacillus thuringiensis that produces a protein which is toxic to insects but
not to humans.

4. Energy Production. Recombinant DNA technology has tremendous scope in energy production.
Through this technology Ii is now possible to bioengineer energy crops or biofuels that grow rapidly
to yield huge biomass that used as fuel or can be processed into oils, alcohols, diesels, or other
energy products. The waste from these can be converted into methane. Genetic engineers are trying
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to transfer gene for cellulase to proper organisms which can be used to convert wastes like sawdust
and cornstalks first to sugar and then to alcohol.
Benefits and Risks of Genetically Modified Organisms
Medicine Industry/ Agriculture

WRAP- UP

Genetic engineering can be done with plants, animals, or bacteria and other very small
organisms. Genetic engineering allows scientists to move desired genes from one plant or animal into
another. Genes can also be moved from an animal to a plant or vice versa. Another name for this is
genetically modified organisms, or GMOs.

The process to create GE foods is different than selective breeding. This involves selecting
plants or animals with desired traits and breeding them. Over time, this results in offspring with those
desired traits.

One of the problems with selective breeding is that it can also result in traits that are not
desired. Genetic engineering allows scientists to select one specific gene to implant. This avoids
introducing other genes with undesirable traits. Genetic engineering also helps speed up the process of
creating new foods with desired traits.

VALUING

In the past century, the recombinant DNA technology was just an imagination that desirable
characteristics can be improved in the living bodies by controlling the expressions of target genes.
However, in recent era, this field has demonstrated unique impacts in bringing advancement in human
life. By virtue of this technology, crucial proteins required for health problems and dietary purposes can
be produced safely, affordably, and sufficiently. This technology has multidisciplinary applications and
potential to deal with important aspects of life, for instance, improving health, enhancing food
resources, and resistance to divergent adverse environmental effects. Particularly in agriculture, the
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genetically modified plants have augmented resistance to harmful agents, enhanced product yield, and
shown increased adaptability for better survival. Moreover, recombinant pharmaceuticals are now being
used confidently and rapidly attaining commercial approvals. Techniques of recombinant DNA
technology, gene therapy, and genetic modifications are also widely used for the purpose of
bioremediation and treating serious diseases. Due to tremendous advancement and broad range of
application in the field of recombinant DNA technology, this review article mainly focuses on its

importance and the possible applications in daily life.

POST-ASSESSMENT

ENRICHMENT ACTIVITIES

Enrichment Activity 1. PAIR ME UP. Choose from the box the corresponding steps or processes
involved in the formation of recombinant DNA as shown in the pictures. Write your answer on the space
below each picture. After which arrange the steps in sequential order.

Outline of recombinant DNA

Enrichment Activity 2. Determine which technologies are most appropriate for which cell types.
TECHNOLOGY CELL TYPE
1. Plants cells
2. Electroporation
3. Biolistics
4. Bacterial cells
5. Mammalian cells

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POST-TEST

Multiple Choice. Read each situation carefully. Choose the letter of the best answer.

Which enzyme is used to cut the piece of the donor DNA?

A. Protoplast
B. DNA ligase
C. Sticky ends
D. Restriction endonuclease
1. Which enzyme is used to cut the piece of the donor DNA?
a. Protoplast b. DNA ligase c. Sticky ends d. Restriction
endonuclease

2. Which genetic engineering process use significant amount of electricity to form pores in the cell
that enables the entry of foreign DNA?
a. Microinjection b. Electroporation c. Protoplast fusion d. Use of particle gun

3. What process wherein the host cell is bombarded with tungsten particles coated with foreign
DNA?
a. Microinjection c. Protoplast fusion
b. Electroporation d. Use of particle gun

4. Which genetic engineering process wherein foreign gene is injected through mild suction in the
host cell?
a. Microinjection c. Protoplast fusion
b. Electroporation d. Use of particle gun

5. What process in which a selected portion of the foreign DNA is inserted into a Plasmid found in
bacteria?
a. Transduction b. Transformation c. Electroporation d. Vectorless gene transfer

6. Which organisms used during transformation process?

a. Vectors b. Plasmid c. Sticky ends d. Transgenic plants

7. What genetic engineering process uses bacteriophages wherein recombinant DNA is

introduced?
a. Transduction b. Transformation c. Electroporation d. Vectorless gene transfer

8. What is the process when a piece of DNA is combined with another DNA from other source?
a. RNA splicing b. Polyadenylation c. DNA transcription d. DNA recombination

9. Which genetic engineering process involves the use of polyethylene glycol for recombination of
genes?
a. Microinjection b. Electroporation c. Protoplast fusion d. Use of particle gun

10. Why genetic markers code important?

a. Pest resistance c. Weather resistance
b. Insect resistance d. Antibiotic resistance

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11. What is the process of making changes on the genetic code of an organism?
a. Civil engineering c. Chemical engineering
b. Genetic engineering d. Biological engineering

12. All of the following processes involve vectors, EXCEPT __.

a. Transduction b. Microinjection c. Transformation d. Bacteriophages

13. All of the following is the importance of plasmid in transformation process EXCEPT ___.
a. Naturally found in bacteria c. It contains a gene sequence
b. It contains a genetic marker d. Composed of tungsten particles

14. What is the correct sequence of the following processes involve in protoplast fusion?
I. Protoplasts fuse
II. Recombinant cell grows new cell wall
III. Segments of the two chromosomes recombine
IV. In solution, protoplasts are treated with polyethylene glycol
V. Bacterial cell walls are enzymatically digested, producing

a. I, III, IV, II, V b. II, III, I, V, IV c. IV, I, V, III, II d. III, IV, II, V, I

15. Is it true that (I) vectors was used during transformation process and (II) electricity also used
during electroporation process?
a. I only b. II only c. Both I and II d. Neither I nor II

16. What bacterium produces proteins rapidly which is use in developing biotherapeutics and
vaccines?
a. Bacillus thuringiensis c. Pseudomonas fluorescens
b. Pseudomonas aeruginosa d. Agrobacterium tumefaciens

17. What is the effect of Agrobacterium tumefaciens to living things?

a. Can cause blindness in humans c. Can cause weight loss in humans
b. Used to introduce DNA to plants d. Used for recombinant DNA in fungi

18. Which bacteria are poisonous to certain pests?

a. Bacillus thuringiensis c. Pseudomonas fluorescens
b. Pseudomonas aeruginosa d. Agrobacterium tumefaciens

19. What is the application of recombinant DNA in the field of agriculture?

a. Maintain good health c. Improved nutritional value
b. To treat human disease d. Developed insect-resistant plant

20. What is being manipulated in genetic engineering?

a. Amino acid b. DNA c. Protein d. RNA

21. What microorganisms used in recombinant DNA?

a. Algae b. Bacteria c. Fungi d. Virus

22. Which is a product of recombinant DNA in plants?

a. Cheese b. Bt corn c. Vaccines d. Gene therapy

23. What recombinant bacterium is called the ice-minus bacterium?

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a. Bacillus thuringiensis c. Pseudomonas fluorescens

b. Pseudomonas syriangae d. Agrobacterium tumefaciens

24. What organism contains genes from other organisms?

a. Macrotrans b. Recomorgi c. Transgenic d. Transplant

25. All of the following are the applications of recombinant DNA in the field of Agriculture, EXCEPT.
a. Virus resistance b. Insect resistance c. Altered Oil content d. Monoclonal antibodies

26. What is the importance of the application of recombinant DNA in the field of food industry?
a. To treat disease c. Process fermented foods
b. Develop vaccines. D. Produce natural insecticides

27. What importance of recombinant DNA is NOT included in the field of agriculture?
a. Bt corn b. Insulin c. Frost crystal d. Weather-guard genes

28. What genetically engineered traits is NOT the application of recombinant DNA to agriculture?
a. Virus resistance c. Herbicide resistance
b. Genetic diseases d. Cold and Drought tolerance

29. Is it true that (I) Recombinant technology is used for processing fermented foods while (II)
Sweetness and color of fruits is also a product of transgenic plant?
a. I only b. II only c. Both I and II d. Neither I nor II

30. How recombinant DNA technology is used in medicine?

I. DNA is put into specially engineered host cell
II. Medicine is given to patient who was prescribed it
III. The medicine produced from DNA is specially packaged
IV. Special DNA sections are stitched together in a plasmid
V. The host cells are grown in batches producing the medicine

a. V, I, III, II, IV b. I, IV, V, II, III c. IV, I, V, III, II d. II, IV, I, V, III

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