General Biology 2: 2 Semester - Module 1
General Biology 2: 2 Semester - Module 1
General Biology 2: 2 Semester - Module 1
Department of Education
Regional Office IX, Zamboanga Peninsula
SHS
GENERAL BIOLOGY 2
2 nd Semester - Module 1
Recombinant DNA
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What's In
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The central dogma of molecular biology explains the flow of genetic information
from genes to protein. It provides a molecular mechanism in understanding how
genotype translates to phenotype and how changing an organismal trait is possible by
altering its genetic makeup. Activity 1 will check your basic knowledge on protein
synthesis and how DNA modification cause changes in the characteristics of
organisms.
Activity 1: DECODE ME
Directions: Use each set of jumbled given letters to reveal the answer for each item.
Use the sentences given as your clue.
___________ DAN 1. Large biomolecule that contains the complete
genetic information for an organism.
___________ INOMA DAIC 2. Building blocks of proteins.
___________ AEELLL 3. It refers to the different forms of genes representing
a certain trait.
___________ UILONTEDCE 4. It is the building block of DNA composed of a five-
carbon sugar, a phosphate group, and a
nitrogenous base.
___________ NEGE 5. A segment of DNA that is capable of storing
information, capable of self-replication, and can
undergo mutation.
___________ RANm 6. It is the molecule that leaves the nucleus during
translation.
___________ EPRLIACTOIN 7. The process wherein DNA molecules are duplicated
during cell division and passed on to each daughter
cell.
___________ NOGEME 8. The entire set of genes for an organism
___________ UNTATIMO 9. A change in the sequence of DNA.
___________ TIONLATARNS 10. The process of synthesizing protein as directed by
the mRNA. It happens in the cytoplasm, near where
the ribosomes stay.
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What's New
1. Write an observation on the color of the rice for the two samples.
Observation: _________________________________________________________________
2. Based on experience and knowledge, what is the common or natural color of the
rice?
_____________________________________________________________________________
The image shows two rice samples of different colors. The difference in the color
is due to the modification in the genetic makeup of the rice through genetic
engineering.
What Is It
Genetic Engineering
Genetic Engineering is a process of altering the genes that are found in all living
organisms. It involves the transfer of genes or parts of DNA from one organism to
another. Organisms whose genes are altered or modified for a specific purpose are
called transgenic organisms.
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How Is DNA Used in Genetic Engineering?
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Adapted from: Introduction to Genetic Engineering and Its Applications. Retrieved from
https://www.teachengineering.org/lessons/view/uoh_genetic_lesson01
Genetically modified organisms are organisms whose genetic material has been
synthetically manipulated in a laboratory through genetic engineering. GMOs refer
broadly to organisms that are produced when selected individual genes are transferred
from a given donor organism into another target organism, typically conferring desired
properties to the new organism. GMOs can include plants, animals, and enzymes.
Some GMOs have been approved by regulatory agencies for commercial
production and consumption, while others are currently undergoing regulatory
evaluation. Practically, all commercial GMOs are engineered to withstand direct
applications of herbicide and/ or to produce an insecticide.
Still, other GMOs are in experimental stages and confined to scientific laboratory
research. According to the United States Department of Agriculture (USDA), by 2012,
93% of soybeans, 94% of cotton, and 88% of corn grown in the U.S. were genetically
modified (Center for Eco genetics and Environmental Health).
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Image Source: From Corgis to Corn: A Brief Look at the Long History of GMO Technology (2015)
Retrieved from http://sitn.hms.harvard.edu/flash/2015/from-corgis-to-corn-a-brief-look-at-the-long-history-of-gmo-technology/
last February 4, 2021
Classical Breeding
o It focuses on the mating of organisms with desirable characteristics.
o May develop new plant varieties in the selection process and seek to achieve
expression of genetic material present within a species.
o It employs processes that occur in nature, i.e., sexual and asexual reproduction.
o The product emphasizes certain characteristics and not new for the species, which
have been present for millennia within the species' genetic potential (Hansen,
2000).
Genetic Engineering
o Done through the insertion of genetic material and must be followed up by gene
insertion.
o The insertion process does not occur in nature; therefore, a gene "gun," a bacterial
"truck," inserts the genetic material into the host plant cells.
This genetic material inserts itself into the chromosomes of the host plant.
Engineers must also insert a "promoter" gene from a virus as part of the package
to make the inserted gene express itself.
o This process alone, involving a gene gun or a similar technique, and a promoter, is
profoundly different from conventional breeding, even if the primary goal is only to
insert genetic material from the same species (Hansen, 2000).
MODIFYING TECHNIQUE
ENHANCED TRAIT (Classical breeding/ Recombinant REASON
DNA technology)
1. Kobe / Wagyu Beef
(Beef with good fat
distribution)
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2. Guapple (Large sized
guava)
3. Human Insulin-
producing bacteria
4. Flavr-Savr (Delayed-
ripening tomatoes)
5. Macapuno trait in
coconuts
Gene Cloning
A process by which large quantities of a specific, desired gene or section of DNA
may be cloned or copied once the desired DNA has been isolated.
What's More
Recombinant DNA
Recombinant DNA technology is a technique that changes the phenotype of an
organism (host) when a genetically altered vector is introduced and integrated into the
genome of the organism. So, the process involves the introduction of a foreign piece of
DNA structure into the genome, which contains our gene of interest. This gene that is
introduced is the recombinant gene, and the technique is called recombinant DNA
technology. Inserting the desired gene into the genome of the host is not as easy as it
sounds. It involves the selection of the desired gene for administration into the host,
followed by a selection of the perfect vector with which the gene has to be integrated
and recombinant DNA formed. This recombinant DNA then has to be introduced into
the host. And at last, it has to be maintained in the host and carried forward to the
offspring (Shinde et.al. 2018).
The primary tools of recombinant DNA technology are bacterial enzymes
called restriction enzymes. Each enzyme recognizes a short, specific nucleotide
sequence in DNA molecules and cuts the molecules' backbones at that sequence. The
result is a set of double-stranded DNA fragments with single-stranded ends, called
"sticky ends." Sticky ends are not really sticky; however, the bases on the sticky ends
form base pairs with the complementary bases on other DNA molecules. Thus, the
sticky ends of DNA fragments can be used to join DNA pieces originating from different
sources.
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Image Source: Recombinant DNA Techniques. Retrieved from http://www.accessexcellence.org
/RC/AB/WYW/wkbooks/SFTS/activity6 last February 4, 2021
Direction: Using the Restriction Enzyme Sequence Cards found in Appendix A and B
in this module, do the following and answer the discussion questions:
1. Cut out the plasmid strips along the dotted lines. Tape the strips together to form a single
long strip. The letters should all be in the same direction when the strips are taped. The
two ends of the strip should then be taped together with the genetic code facing out to
form a circular plasmid.
2. Cut out the DNA base sequence strips, and tape them together to form one long strip. The
pieces must be taped together in the order indicated at the bottom of each strip.
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3. Next, cut out the restriction enzyme cards. Take note that the enzyme cards illustrate a
short DNA sequence that shows the sequence that each particular enzyme cuts.
4. Compare the sequence of base pairs on an enzyme card with the sequences of the
plasmid base pairs. If you find the same sequence of pairs on both the enzyme card and
the plasmid strip, it should be marked as the location on the plasmid with a pencil, and
write the enzyme number in the marked area. Do this for each enzyme card. The enzyme
sequences may not have a corresponding sequence on the plasmid, and that some
enzyme sequences may have more than one corresponding sequence on the plasmid.
5. Once all corresponding enzyme sequences are identified on the plasmid, identify those
enzymes which cut the plasmid once and only once. Discard any enzymes that cut the
plasmid in the shaded plasmid replication sequence. Record your findings.
6. Compare the enzymes you listed against the cell DNA strip. Find any enzymes that will
make two cuts in the DNA, one above the shaded insulin gene sequence and one below
the shaded insulin gene sequence. Mark the areas on the DNA strip that each enzyme will
cut.
7. Select one enzyme to use to make the cuts. The goal is to cut the DNA strand as closely
as possible to the insulin gene sequence without cutting into the gene sequence. Have the
students make cuts on both the plasmid and the DNA strips. They should make the cuts
in the staggered fashion indicated by the black line on the enzyme card.
8. Tape the sticky ends (the staggered ends) of the plasmid to the sticky ends of the insulin
gene to create their recombinant DNA.
A. Discussion Questions:
1. Why was it important to find an enzyme that would cut the plasmid at only one
site?
__________________________________________________________________________________
__________________________________________________________________________________
2. What could happen if the plasmid were cut at more than one site?
__________________________________________________________________________________
__________________________________________________________________________________
3. Why was it important to discard any enzymes that cut the plasmid at the replication
site?
____________________________________________________________________________________
_________________________________________________________________________________
4. Why might it be important to cut the DNA strand as closely to the desired gene as
possible? In this activity, you incorporated an insulin gene into the plasmid. How
will the new plasmid DNA be used to produce insulin?
____________________________________________________________________________________
________________________________________________________________________
Sequence Steps
1. Reintroduce donor gene into donor cells.
2. Modify donor gene.
3. Identify donor gene of interest by using crosses.
4. Clone donor gene of interest in bacterium.
5. Characterize donor gene in bacterial system.
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1. Production of Transgenic Plants
By utilizing the tools and techniques of genetic engineering, it is possible to
produce transgenic plants or genetically modified plants. Many transgenic plants have
been developed with better qualities like resistance to herbicides, insects, or viruses or
with the expression of male sterility, etc.
2. Production of Transgenic Animals
By the use of rec DNA technology, desired genes can be inserted into the animal
so as to produce the transgenic animal. The method of rec DNA technology aids the
animal breeders to increase the speed and range of selective breeding in the case of
animals. It helps for the production of better farm animals to ensure more commercial
benefits. Another commercially important use of transgenic animals is the production
of specific proteins and pharmaceutical compounds. Transgenic animals also
contribute to studying the gene functions in different animal species. Biotechnologists
have successfully produced transgenic pigs, sheep, rats, and cattle.
3. Production of Hormones
By the advent of techniques of rec DNA technology, bacterial cells like E.coli are
utilized for the production of different fine chemicals like insulin, somatostatin,
somatotropin, and endorphin. Human Insulin Hormone, i.e., Humulin, is the first
therapeutic product that was produced by the application of rec DNA technology.
4. Production of Vaccines
Vaccines are the chemical preparations containing a pathogen in an attenuated (or
weakened) or inactive state that may be given to human beings or animals to confer
immunity to infection. A number of vaccines have been synthesized biologically
through recDNA technology; these vaccines are effective against numerous serious
diseases caused by bacteria, viruses, or protozoa. These include vaccines for polio,
malaria, cholera, hepatitis, rabies, smallpox, etc. The generation of DNA vaccines has
revolutionized the approach to the treatment of infectious diseases. DNA-vaccine is the
preparation that contains a gene encoding an immunogenic protein from the
concerned pathogen.
5. Biosynthesis of Interferon
Interferons are the glycoproteins that are produced in very minute amounts by
the virus-infected cells. Interferons have antiviral and even anti-cancerous properties.
By the recDNA technology method, the gene of human fibroblasts (which produce
interferon's in human beings) is inserted into the bacterial plasmid. These genetically
engineered bacteria are cloned and cultured so that the gene is expressed and the
interferons are produced in relatively high quantities. This interferon, so produced, is
then extracted and purified.
6. Production of Antibiotics
Antibiotics produced by microorganisms are very effective against different viral,
bacterial, or protozoan diseases. Some important antibiotics are tetracycline,
penicillin, streptomycin, novobiocin, bacitracin, etc. The recDNA technology helps in
increasing the production of antibiotics by improving the microbial strains through
modification of genetic characteristics.
7. Production of Commercially Important Chemicals
Various commercially important chemicals can be produced more efficiently by
utilizing the methods of rec DNA technology. A few of them are the alcohols and
alcoholic beverages obtained through fermentation, organic acids like citric acid, acetic
acid, etc., and vitamins produced by microorganisms.
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8. Application in Enzyme Engineering
As we know that the enzymes are encoded by genes, so if there are changes in a
gene, then definitely the enzyme structure also changes. Enzyme engineering utilizes
the same fact and can be explained as the modification of an enzyme structure by
inducing alterations in the genes which encode for that particular enzyme.
9. Prevention and Diagnosis of Diseases
Genetic engineering methods and techniques have greatly solved the problem of
conventional methods for the diagnosis of diseases. It also provides methods for the
prevention of a number of diseases like AIDS, cholera, etc. Monoclonal antibodies are
useful tools for disease diagnosis. Monoclonal antibodies are produced by using the
technique called hybridoma technology.
10. Gene Therapy
Gene therapy is undoubtedly the most beneficial area of genetic engineering for
human beings. It involves the delivery of specific genes into the human body to correct
the diseases. Thus, it is the treatment of diseases by transfer and expression of a gene
into the patients' cells so as to ensure the restoration of a normal cellular activity.
What I Can Do
Activity 7: MATCHY MATCHY
Direction: Match the purpose to the components found in the box below.
Antibiotic Multiple Cloning Site Promoter
Resistance Gene
DNA Inserted Gene Sequence Multiple Cloning Site
1. Allows the controlled expression of the desired gene in the
presence of an inducing agent (e.g., beta-galactosidase; heat
treatment (~65°"C)
2. DNA sequence or portion for the insertion of the desired gene.
This section may contain sequences that will be cut by
specific restriction endonucleases ( cuts within the molecule)
3. Successful insertion of a gene allows the expression of its
protein product. This usually provides a specific trait to the
"transformed" bacteria.
4. Provides a way to screen a population of bacteria for those
that took up the plasmid. For example, if an ampicillin
resistance gene is encoded in the plasmid, then only bacteria
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that took up the plasmid will grow on media with ampicillin.
5. The gene for Green Fluorescent Protein is placed within the
expression plasmid; bacteria transformed with this plasmid
will produce a protein (GFP) that will allow the bacterial
cells/colonies to glow green in the dark.
Assessment
Direction: Encircle the letter of the best answer.
1. What carries a gene from one organism into a bacteria cell?
A. a plasmid C. a restriction enzyme
B. an electrophoresis gel D. polymerase chain reaction
2. What is a genetically modified organism (GMO)?
A. a plant with certain genes removed
B. an organism with an artificially altered genome
C. a hybrid organism
D. any agricultural organism produced by breeding or biotechnology
3. What is the role of Agrobacterium tumefaciens in the production of transgenic
plants?
A. Genes from A. tumefaciens are inserted into plant DNA to give the plant
different traits.
B. Transgenic plants have been given resistance to the pest A. tumefaciens.
C. A. tumefaciens is used as a vector to move genes into plant cells.
D. Plant genes are incorporated into the genome of Agrobacterium tumefaciens.
4. What is the most challenging issue facing genome sequencing?
A. the inability to develop fast and accurate sequencing techniques
B. the ethics of using information from genomes at the individual level
C. the availability and stability of DNA
D. all of the above
5. Genomics can be used in agriculture to:
A. generate new hybrid strains C. improve yield
B. improve disease resistance D. all of the above
6. Can scientists predict with certainty where an inserted gene will go on a plant
chromosome?
A. With modern genetic techniques, scientists can insert genes precisely.
B. Genes are inserted on the proper chromosome, but there is no control on
where it goes on the chromosome.
C. Scientists have a general idea of where the gene will go and what it will do to
the plant.
D. It's just a shot in the dark.
7. Can genes escape from genetically modified crops and jump to other plants?
A. Yes, and often do.
B. Only to some crops, but those crops aren't genetically modified.
C. Only during rare climatic conditions.
D. No, genes cannot move from species to species without human intervention.
8. Which of the following steps is NOT essential in producing recombinant DNA?
A. Cut out a piece of DNA from a DNA molecule.
B. Insert a piece of DNA from one organism into the DNA of another organism.
C. Use a restriction enzyme to cut DNA and form sticky ends.
D. Read the sequences of bases in a piece of DNA.
9. To produce transgenic bacteria that make insulin, which of the following steps did
scientists have to take first?
A. Insert the human insulin gene into a plasmid.
B. Extract the insulin from the bacterial culture.
C. Use a restriction enzyme to cut out the insulin gene from human DNA.
D. Transform bacteria with the recombinant plasmid.
10. DNA from a human has been inserted into a bacterial plasmid and reinserted back
into the bacterium. The bacterium now contains both human DNA and bacterial
DNA. The bacterium is now considered as a/an __________.
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A. mutation B. PCR C. clone D. transgenic organism
Additional Activities
Activity 8: Infographics
Direction: Create an infographic showing the Pros and Cons of Genetic
Engineering. Use the given rubric as a guide.
3 POINTS: EXCEEDS 2 POINTS: MEETS 1 POINT:
EXPECTATIONS EXPECTATIONS NEEDS WORK
Topic/Purpose The topic/purpose of the The topic/purpose was The topic/purpose of
infographic was clear and somewhat broad and did not the infographic was
concise. allow the viewer to not clear and concise.
understand the intent.
Data of the infographic was Data of the infographic was Data of the infographic
accurate and relevant to the somewhat accurate and was not accurate and
Data topic relevant to the topic. was not relevant to the
topic.
The infographic had a great The graphics were somewhat The graphics had
layout, with applicable applicable to the infographic, nothing to do with the
Layout graphics. creating an average layout. topic and had a poor
layout. There was an
overload of text.
The font was legible, and the The font was somewhat The font was not
color scheme enhanced the legible, and the color scheme legible, and the color
Color/Font infographic. didn't affect the infographic. scheme detracted from
the infographic.
Citations for the infographic's Citations for some of the No citations of the
Sourcing sources were included. sources used were included. infographic's sources
were included.
Source: https://www.uen.org/rubric/previewRubric.html?id=30103
1. Cut out strips along dotted lines. 1. Cut out strips along dotted lines.
2. Tape together top to bottom in any order. 2. Tape together top to bottom in
numeral order
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Shaded region = insulin gene site
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References
Applications of Genetic Engineering. Retrieved from
https://batch.libretexts.org/print/url=https://bio.libretexts.org/Bookshelves/Microbiology/Book%3A_Microbio
logy_(Boundless)/7%3A_Microbial_Genetics/7.23%3A_Genetic_Engineering_Products/7.23B%3A__Applications_
of_Genetic_Engineering.pdf last February 2, 2021.
Fast Facts about Genetically Modified Organisms. Retrieved from
https://depts.washington.edu/ceeh/downloads/FastFacts_GMOs_FINAL.pdf
Genetic Engineering Flow Chart. Retrieved from
https://www.teachengineering.org/content/uoh_/lessons/uoh_genetic/uoh_genetic_lesson01_flowchart_v2_ted
l_dwc.pdf last February 2, 2021.
Genetically Modified Food1 Keith R. Schneider, Renée Goodrich Schneider, and Susanna Richardson Retrieved from
https://edis.ifas.ufl.edu/pdffiles/FS/FS08400.pdf
Department of Education- Division of Cagayan de Oro City) (2020). General Biology 2- Quarter 1 - Module 1: Genetics
Hansen (2000) GENETIC ENGINEERING IS NOT AN EXTENSION OF CONVENTIONAL PLANT BREEDING; How genetic
engineering differs from conventional breeding, hybridization, wide crosses, and horizontal gene transfer.
Retrieved from https://advocacy.consumerreports.org/wp-content/uploads/2013/02/Wide-Crosses.pdf
Shinde, at. al (2018) Recombinant DNA Technology and its Applications: A Review: International Journal of MediPharm
Research, Vol.04, No.02, pp 79-88, 2018
The Commission on Higher Education. Teaching Guide for Senior High School General Biology 2
Department of Education Central Office. Most Essential Learning Competencies ( MELCs). 2020.
https://bio.libretexts.org/Bookshelves/Introductory_and_General_Biology/Book%3A_Concepts_in_Biology_(OpenStax)
/10%3A_Biotechnology/10.E%3A_Biotechnology_(Exercises)
https://www.purdue.edu/uns/html4ever/0007.Goldsbrough.cropquiz.html
https://www.cwcboe.org/cms/lib/NJ01001185/Centricity/Domain/143/Chapter%2012%20-
%20Genetic%20Engineering/Practice%20test/practice%20test%20answers.pdf
Recombinant DNA Techniques. Retrieved from
http://www.accessexcellence.org/RC/AB/WYW/wkbooks/SFTS/activity6 last February 4, 2021
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