Module 1

1. It is a double-stranded, self-replicating, circular DNA molecule present inbacteria which is widely used as a gene
cloning vector.

A. Cosmid B. Plasmid C. Phagemid D. Genome

2. These enzymes are called molecular scissors which is essential in making internal cuts in a DNA molecule or vector
at specific sites.

A. Restriction enzymes B. Restriction proteins

C. Polymerase D. All of these

3. What do we call the new molecule after the process of ligation wherein the vector plasmid joined with the gene of
interest?

A. Gene of interest B. Recombinant DNA

C. Chimeric DNA D. All of these

4. During transfer of the recombinant plasmid into a host cell which is commonly a bacterium, the recombinant
bacterium will undergo cloning. Which of the following is involve in gene cloning?

A. Replication B. Transcription

C. Translation D. Duplication

5. Bacterial cells as host organisms may express the gene to make protein products such as insulin and growth
hormones. Which of the following processes are involved?

A. Replication and transcription B. Transcription and translation

C. Transcription and transformation D. Replication and transformation

6. In plant genetic engineering, which of the following acts as vector?

A. Agrobacterium tumefaciens B. Gene of interest

C. Recipient plant cell D. Ti-plasmid

7. Which of the following describes recombinant DNA technology?

A. Mating of organisms with desirable qualities

B. Insertion of genes into cells that makes the cells into “factories” to make products

C. Enhancing or disrupting the traits of a target organism either mating or molecular technique

D. All of the above

8. Which of the following illustrates classical breeding?

A. A farmer choose a breed of cow of greater milk production

B. The use of bacteria in order to produce human insulin

C. The insertion of cloned genes to plant cells

D. All of the above

9. Which of the following is/are example of genetic engineering?

A. A farmer choose a breed of cow for of greater milk production

B. The use of bacteria in order to produce human insulin

C. The insertion of cloned genes to plant cells

D. Cross pollination of squash flowers

10. Which of the following is an example of genetically modified plant?

A. Seedless grapes B. Guapple (larged-sized guava)

C. Rice with beta-carotene D. All of these

11. Being pest resistance is one of the traits that are being introduced to plants like corn and eggplant with the
insertion of Bt-toxin gene to plant cell. What method is use when a gene is inserted to plants?

A. Biolistic B. Electroporation

C. Plasmid insertion by heat shock treatment D. All of these

12. Why recombinant DNA is very useful in improving our health condition?

A. Human insulin can be reproduced by bacteria

B. Vaccine can be reproduced by fungi or bacteria

C. Human growth hormones can be obtained from E.coli

D. All of the above

13. A biotechnologist wants to enhance the size of duhat fruit because research shows that it can lower blood sugar
but is very expensive in the market. What could be the possible source organism which contains the gene that is
responsible for the development of large fruits?

A. Lanka B. Chico C. Strawberry D. Mango

14. Mario wants a hybrid or mestizo breed of pigs to raise that’s why he chooses to mate his native pig (pure breed
black) to a white pig (pure breed white). What type of genetic engineering is shown in the situation?

A. Classical breeding B. Artificial selection

C. Recombinant DNA D. All of these

15. Why genetic engineering is beneficial to the society?

A. It improves crop varieties to meet the demand of increasing human population

B. production of human proteins using bacteria is a great leap in the field of medicine

C. Transgenic plants and transgenic animals play a great role in our economic

D. All of the above

Module 2
1. Which organism first dominated the Earth?

A. Dinosaurs B. Fishes C. Plants D. Bacteria

2. The Mesozoic era is called the Age of reptiles, how about the Cenozoic era?

A. Age of mammals B. Age of birds

C. Age of humans D. Age of Technology

3. What is the longest part of Earth’s history which covers 88% of the Earth’s history?

A. Precambrian B. Paleozoic C. Mesozoic D. Cenozoic

4. The geologic time scale is subdivided into four groups. What is the correct sequence if you will arrange them from
the largest to the smallest?

A. Eon, period, epoch, era B. Eon, era, epoch, period

C. Eon, era, period, epoch D. Era, eon, period, epoch

5. Coronacollina acula is a fossil that represent the oldest animal with hard body parts that was discovered recently in
South Australia. Scientist believed that this animal existed during ___________.

A. Precambrian B. Paleozoi C. Mesozoic D. Cenozoic

6. Which era were the first land plants formed?

A. Precambrian B. Paleozoic C. Mesozoic D. Cenozoic

7. Which event occurred during the Mesozoic era?

A. Pangea formed B. Asteroid killed the dinosaurs

C. Rocky mountain formed D. Pleistocene ice age began

8. Approximately, how many mass extinction events occurred throughout the evolutionary history of animals?

A. 3 B. 4 C. 5 D. more than 5

9. Until recent discoveries suggested otherwise, animals existing before the Cambrian period were believed to be
__________’

A. small and ocean dwelling B. small and nonmotile

C. small and soft- bodied D. small and radially symmetrical

10. Which scientific concepts did Charles Darwin and Alfred Wallace independently

discover?

A. Mutation B. Genetic Drift C. Natural selection D. Recombination

11. Which of the following evolutionary forces can introduce new genetic variation into a population?

A. Natural selection and genetic drift B. Mutation and gene flow

C. Mutation and genetic drift D. natural selection and non-random mating

12. Which of the following mechanism of change was brought by the exchange of genes during the process of meiosis
that leads to the formation of new gametes that are different form its parent?

A. Mutation B. Genetic Drift

C. Natural selection D. Recombination

13. Which of the following situations will lead to natural selection?

A. The seeds of two plants land near each other and one grows longer than the other

B. Two types of fish eat the same kind of food, and one is better able to gather food than the other.

C. Male lions compete for the right to mate with females, with only one possible winner

D. All of the above

14. When male lions reach sexual maturity, they leave their group in search of a new pride. This can alter the allele
frequencies of the population through which of the following mechanisms?

A. Natural selection B. Artificial selection

C. Recombinant DNA D. All of these

15. Which of the following statement is TRUE about the different mechanism of evolution?

A. Natural selection works by selecting alleles that confer beneficial traits orbehavior.

B. Mutations introduce new alleles into a population.

C. Allele frequencies can change as a result of gene flow

D. All of the above

Module 3
1. Which of the following must exist in a population before natural selection can act upon that population?

A. Genetic variation among individuals B. Variation among individuals caused by environmental factors

C. Sexual reproduction D. Three of the responses are correct.

2. Which of the following does not contribute to the study of evolution?

A. Population genetics B. Inheritance of acquired characteristics

C. Fossil record D. Comparative embryology

3. Catastrophism, meaning the regular occurrence of geological or meteorological disturbances (catastrophes), was
Cuvier's attempt to explain what existence?

A. Evolution. C. Uniformitarianism.

B. The fossil record. D. The origin of new species.

4. Which of the following represents an idea that Darwin learned from the writings of Thomas Malthus?

A. Technological innovation in agricultural practices will permit exponential growth of the human population into the
foreseeable future.

B. Populations tend to increase at a faster rate than their food supply normally allows.

C. Earth changed over the years through a series of catastrophic upheavals.

D. The environment is responsible for natural selection.

5. In the mid-1900s, the Soviet geneticist Lysenko believed that his winter wheat plants, exposed to ever-colder
temperatures, would eventually give rise to ever more cold-tolerant winter wheat. Lysenko's attempts in this regard
were most in agreement on which of the following scientist?

A. Cuvier B. Hutton C. Lamarck D. Darwin

6. Which of the following ideas is not included in Darwin’s theory?

A. All organisms that have ever existed arose through evolutionary modifications of ancestral species.

B. Natural selection drives some evolutionary change.

C. Natural selection preserves favorable traits.

D. Natural selection eliminates adaptive traits.

7. Which of the following statements is not compatible with Darwin’s theory?

A. All organisms have arisen by descent with modification.

B. Evolution has altered and diversified ancestral species.

C. Evolution occurs in individuals rather than in groups.

D. Natural selection eliminates unsuccessful variations.

8. Which of the following statements about biological species is(are) correct?

I. Biological species is a group of individuals whose members interbreed with one another.

II. Biological species are the model used for grouping extinct forms of life.

III. Members of biological species produce viable, fertile offsprings.

A. I only B. II only C. I and III D. II and III

9. Which of the following is not an isolating mechanisms that prevent fertilization and formation of zygote?

A. Temporal isolation B. Hybrid breakdown

C. Gametic isolation D. Ecological isolation

10. Which of the following is the occurrence of abrupt genetic change cause reproductive isolation between groups of
individuals?

A. Allopatric speciation B. Convergent speciation

C. Divergent speciation D. Sympatric speciation

11. Which of the following speciation occurs when populations are separated by a geographic barrier?

A. Allopatric speciation B. Convergent speciation

C. Divergent speciation D.Sympatric speciation

12. Which of the following is the abrupt change in the environment over a geographic border and strong disruptive
selection affects gene flow between neighboring populations?

A. Allopatric speciation B. Convergent speciation

C. Divergent speciation D. Sympatric speciation

13. Which of the following statements about natural selection is CORRECT?

A. Adaptations beneficial in one habitat should generally be beneficial in all other habitats as well.

B. Adaptations beneficial at one time should generally be beneficial during all other times as well.

C. Different species that occupy the same habitat will adapt to that habitat by undergoing the same genetic changes.

D. Well-adapted individuals leave more offspring, and thus contribute more to the next generation's gene pool, than do
poorly adapted individuals.

14. Which of the following factors BEST explains why the human population has grown so rapidly over the last 1000
years?

A. Humans have decreased their reliance on natural resources

B. Humans have increased the amount of resources available on Earth.

C. Humans have increased the carrying capacity of the biosphere for the population.

D. Humans have developed physical characteristics that increase their competitive advantage.

15. Which of the following branches of biology deals with naming and classifying organisms?

A. Binomial Nomenclature B. Evolution

C. Genetics D. Taxonomy
Module 4
1. DNA of the life-forms on Earth are almost universal and seem to be templated from one original source, this line of
inferring can be based from what evidence of evolution?

A. Fossil record B. Comparative Anatomy

C. Molecular Biology D. Embryology

2. Exactly different organisms such fish and humans may have resemblances during the early stage of their
development and will lose some key likenesses as they grow in later stage of development, which among the
following is the line of evidence and field of study that underlie the said concept for evolution?

A. Fossil record B. Comparative Anatomy

C Molecular Biology D. Embryology

3. Preserved bodies of cave dwelling man from cold and icy places suggested that humans look different in the past,
what evidence of evolution is used?

A. Fossil record B. Comparative Anatomy

C. Molecular Biology D. Embryology

4. Identify what evidence of evolution is used: vertebrate animals such as humans, chickens and fish have gill slits and
tails during their embryonic stage.

A. Fossil record B. Comparative Anatomy

C. Molecular Biology D. Embryology

5. Boa constrictors, a kind of snake has a remnant of what appears to be rudimentary hind legs, what type of body
structure are these?
A. Analogous structures B. Homologous structures

C. Embryonic structures D. Vestigial structure

6. These body structures indicate that organisms descended from a close common ancestor.

A. Analogous structures B. Homologous structures

C. Embryonic structures D. Vestigial structures

7. Fox and polar bears which are distant relatives both developed white colored fur to adapt to the snowy
environment where they habituate. These body structures are identified as:

A. Analogous structures B. Homologous structures

C. Embryonic structures D. Vestigial structures

8. This is the study of body structures of organisms to compare and infer evolutionary links.

A. Fossil record B. Comparative Anatomy

C. Embryology D. Biogeography

9. Which among the following best explains the fossil records as an evidence of evolution?

A. Recorded events from the past indicates that the Earth was once filled with water.

B. Fossils suggest that the Earth is not the same as it is today, for instance there were once a huge massive
interconnected land termed as Pangaea.

C. Fossils recorded the history of life on Earth and indicates that ancient life forms were different from modern day
species.

D. Recorded activities of animals from prehistoric times suggest that people came from monkeys.

10. How will you differentiate analogous structures from homologous structures as evidence of evolution?

A. Analogous structures indicate common ancestry while homologous structures suggest evolution because of same
environmental factors.

B. Analogous structures are structures which indicates that organisms might developed structures with same function as
needs arise while homologous structures showed pattern of common descent from different body structures of closely
related specie.

C. Both analogous and homologous structures are body structures used by researchers to study the evolutionary
development of organisms while vestigial organisms showed different result.

D. B and C

11. What evidence of evolution is portrayed by the unique species on islands which are usually isolated from another
mainland?

A. Fossil record B. Comparative Anatomy

C. Embryology D. Biogeography

12. Mapping of DNA allows scientist to compare the genes of organisms from the past and organisms present today,
the evidence of evolution used is:

A. Fossil record B. Molecular Biology

C. Analogous structures D. Comparative Anatomy

13. Which is NOT true about fossils?

A. They suggest that life has a history.

B. They are proofs of life in the past.

C. The older the fossil, the less it looks like modern day species.

D. They seem precisely like modern-day species, regardless of their age

14. How will you use biogeography as an evidence to infer evolutionary relationship?
A. Organisms living closer at each other and sharing the same niche are also closely related to each other.

B. The unique characteristics of the organisms living on a secluded area is indicative of their ancestry and speciation.

C. Biogeography revealed that organisms with similar developmental pattern even if found at different places might once
live together before they were separated due to natural events or forces.

D. All of the above

15. The following are the pieces of evidence of evolution that may be used to infer the evolutionary relationship
between organisms, EXCEPT _________________.

A. Comparative Anatomy B. Fossil record

C. Molecular Bonds D. Embryology

Module 5
1. Which of the following refers to the two-name classification system used to organize living things?

A. Biodiversity B. binomial nomenclature

C. phylogeny D. systematics

2. What are the main levels of taxa in order?

A. domain, kingdom, phylum, class, order, family, genus

B. kingdom, phylum, class, order, family, genus, species

C. domain, kingdom, phylum, class, order, genus, family, species

D. domain, kingdom, phylum, class, order, family, genus, species

3. Which of the following scientific name is correctly written?

A. Musca domestica B. Musca domestica

C. Musca Domestica D. Musca domestica

4. What is defined as the study of the diversification of living forms, both past and present, and the relationships
among living things through time?

A. Phonetics B. systematics

C. systemics D. taxonomy

5. How important is DNA barcoding in systematics? DNA barcoding helps determine _____________

A. morphological similarities B. evolutionary relationships

C. physiological difference s D. physical traits

6. Which of the following is NOT part of a taxonomist role?

A. Assigning a scientific name.

B. Classifying ornamental plants.

C. Identifying an unknown flowering plant.

D. Explaining the relatedness of plants in different locations.

7. To which category will two organisms with different correlated morphological characters belong to?

A. one biological species B. two biological species

C. one taxonomic species D. two taxonomic species

8. What do you call the type of classification that reflects the evolutionary inter relationships of organisms?

A. Phylogenetic classification B. Artificial classification

C. Natural classification D. Numerical classification

9. What is the characteristic of a rooted phylogenetic tree? Phylogenetic tree is one ________________.

A. that extends back to the origin of life on earth.

B. at whose base is located the common ancestor of all taxa depicted on that tree.

B. that illustrates the rampant gene swapping that occurred early in life’s history.

C. that indicates our uncertainty about the evolutionary relationships of the taxa depicted in the tree.

D. That hides the evolutionary relationships among various biological

species.

10. Which of the following anatomical tree part is most analogous to two species with a common ancestor?

A. the trunk

B. a node where two twigs diverge

C. a twig that branches with time

D. a single twig that gets longer with time

11.Which of the following is a characteristic of the best classification system? A classification system that __________.

A. reflects evolutionary history.

B. conforms to traditional, Linnaean taxonomic practices.

C. unites organisms that possess similar morphologies.

D. corroborates the classification scheme in use at the time of CharlesDarwin.

12. Which level of classification within a lineage has the largest number of shared derived characters?

A. Kingdom B. Class C. Domain D. Family

13-15. Use the figure below to answer questions 13-15.

13. Which number represents the common ancestor for both species C and E?

A. 1 B. 2 C. 3 D. 4

14. Which species are closely related?

A. A and B B. B and C C. C and D D.B and D

15. Which species are extinct?

A. A and E B. B and C C. C and D D.B and D

Geologic time scale – yung binigay ni maam na papel

Early Scientists who contributed in developing evolutionary thought.
Carolus Linnaeus - binomial nomenclature.
- a Swedish botanist who lived during the 1700s. He is known as the “father of taxonomy.”
 - classification system - Systema Naturae (“System of Nature”). - domain, kingdom, phylum, class, order,
family, genus, species

Thomas Malthus - He observed that an increase in a nation's food production improved the wellbeing of the
populace…

- "Malthusian trap" or the "Malthusian spectre" - humans had a propensity to utilize

abundance for population growth rather than for maintaining a high standard of
living.

- Malthusian catastrophe - Populations had a tendency to grow until the lower class suffered
hardship, want and greater susceptibility to famine and disease

Georges Cuvie - • Father of Paleontology

• Theory of Catastrophism = boundaries represent floods, droughts, etc. that destroyed many species
living at that time

• According to him, fossils are remains of extinct life forms

James Hutton - • Theory of Gradualism

- • Proposed that the Earth was shaped by geological forces occurring over very long periods of time,
and is MILLIONS not THOUSANDS of years old.

Charles Lyell - • Principles of Geology

- "uniformitarian

Jean Baptiste Lamarck - One of first scientists to recognize that living things changed over time and that all species were
descended from other species.

- Lamarckism

- 1809- Published his ideas about “Inheritance of Acquired Characteristics”

Charles Darwin

- His voyage and his observations led him to write ‘The Origin of Species

- •He noticed plants and animals were different from those he knew in Europe

• He wrote thousands of pages of observations and collected vast number of Specimens.

• He spent a month observing life on the Galapagos Islands and realized that each island has different rainfall and
vegetation and its own unique assortment of plant and animal species.

• He collected 14 species of finches and hypothesized that the Galapagos had be colonized by organisms from the
mainland that had then diversified on the various.

• In 1859 , his book On the Origin of Species by Means of Natural Selection was published. It presented evidence and
proposed a mechanism for evolution that he called NATURAL SELECTION.

• It took Darwin years to form his theory of evolution by natural selection. His reasoning went like this:

1.Like Lamarck, Darwin assumed that species can change over time. The fossils he found helped convince him of that.
2. From Lyell, Darwin saw that Earth and its life were very old. Thus, there had been enough time for evolution to
produce the great diversity of life Darwin had observed.

3. From Malthus, Darwin knew that populations could grow faster than their resources. This “overproduction of
offspring” led to a “struggle for existence,” in Darwin’s words.

4. From artificial selection, Darwin knew that some offspring have variations that occur by chance, and that can be
inherited. In nature, offspring with certain variations might be more likely to survive the “struggle for existence” and
reproduce. If so, they would pass their favorable variations to their offspring.

5. Darwin coined the term fitness to refer to an organism’s relative ability to survive and produce fertile offspring.
Nature selects the variations that are most useful. Therefore, he called this type of selection natural selection.

6. Darwin knew artificial selection could change domestic species over time. He inferred that natural selection could also
change species over time. In fact, he thought that if a species changed enough, it might evolve into a new species

• Reproductive Isolating Mechanism

- The mechanisms of reproductive isolation are a collection of evolutionary mechanisms, behaviors
and physiological processes critical for speciation. They prevent members of different species from
producing offspring, or ensure that any offspring are sterile

1. Pre-zygotic isolation mechanisms

- prevent fertilization and zygote formation.

- happens before fertilization occurs between gametes.

1.1 Geographic or ecological or habitat isolation

- The two species live in different habitats and will not encounter one another: each is isolated from the other
species.

•1.2 Temporal or seasonal isolation

- different groups may not be reproductively mature. For example, two populations of plants may produce
flowers in different seasons, making mating between the populations impossible.

•1.3 Behavioral isolation

- patterns of courtship is different. For example, eastern & western meadowlark songs differ.

•1.4 Mechanical isolation

- differences in reproductive organs prevent successful interbreeding. Mechanical isolation occurs when mating
is physically impossible

•1.5 Gametic isolation

- incompatibilities between egg and sperm prevent fertilization

2. Post-zygotic isolation mechanisms

- - allow fertilization but nonviable or weak or sterile hybrids are formed. In these cases, the zygote
formed is called a hybrid. However, even after a hybrid zygote forms, reproduction may
still not be successful.

2.1 Hybrid inviability

- fertilized egg fails to develop past the early embryonic stages. For example, when tigers and leopards are
crossed, the zygote begins to develop but the pregnancy ends in miscarriage

or stillborn.

2.2 Hybrid sterility

 -their hybrids are sterile because gonads develop abnormally or there is abnormal segregation of chromosomes
during meiosis. A horse and a donkey may produce a hybrid offspring, a mule. Mules are sterile

2.3 Hybrid breakdown

- F1 hybrids are normal, vigorous and viable, but F2 contains many weak or sterile individuals

CONCRETE EVIDENCE OF EVOLUTION AND HOW TO INFER USING ITS EVOLUTIONARY

RELATIONSHIP
A. Fossils

-Fossils are preserved remnants of once living organisms trapped in rocks, tarpits, frozen in ice or embedded in
amber.

B. Embryology

-An embryo is an organism in its initial phases of development. While embryology is the study of the
development of the anatomy of an organism to its adult form also gives evidence of relatedness
between now widely different groups of organisms

C. Anatomical Evidence

-In comparing the anatomy and the development of organisms, it shows a unity of plan among those that are
closely related. The more body structures that two species have in common, the more closely they are related. It adopts
the idea of “descent from a common ancestor”. Similar structures in different species irrespective of their functions are
called homologous structures. Homology seems to indicate descent from common ancestor. The limb skeletons of
vertebrates are homologous structures.

D. Biochemical

- Living organisms shared numerous related biochemical molecules, such as DNA, ATP, amino acids, and enzymes.
This finding supports descent from a common ancestor.

1. DNA/Nucleic acids

2. Proteins

E. Biogeography

-Biogeography is the study of the geographic distributions of organisms. Darwin’s trip around South America
allowed him to observe the diversity of organisms in different areas and the resemblance of
such species of birds and tortoises in an island to nearby mainland.
Structural and developmental characteristics and relatedness of DNA sequences
Anatomy and embryology

- Anatomical features shared between organisms (including ones that are visible only during embryonic
development) can indicate a shared evolutionary ancestry.

- It is said that physical characteristics shared due to evolutionary history (a common ancestor) are homologous.
To give one example, the forelimbs of whales, humans, and birds are homologous structures.
Not all physical traits that appear identical are indicators of shared ancestors. Instead, some physical
similarities are analogous: in different species, they developed independently because the
organisms lived in similar environments or encountered similar selective pressures. A duck and a platypus
are similar in that they both lay eggs, however, the egg laying capability likely developed
independently rather than from a common ancestor.

Molecular biology

-Structural homologies, similarities may reflect shared evolutionary ancestry between biological molecules.
Similarities and variations in various species between the "same" gene (that is, a pair of
homologous genes) will help us decide how closely the organisms are related.

DNA evidence for evolutionary relationships

- All living organisms share the same genetic material (DNA), identical genetic codes, and the same basic gene
expression mechanism at the most basic level (transcription and translation). The sequences of
associated (or homologous) genes are also contrasted by biologists. If the "same" gene is
found in two animals, it is because they inherited it from a shared ancestor

Taxonomy vs. Systematics

-Systematics is the study of the diversification of life forms over time, both past and present, and their
relationships between other species. On the other hand, taxonomy is the science of organizing
and categorizing living organisms into classes called taxa.

-Both a systematist and a taxonomist provide scientific names; give detailed descriptions of organisms; collects
and keeps volumes of specimens; offer classifications for the organisms by constructing
identification keys and data on their occurrence and distribution. However, it is the systematist that
investigates on evolutionary histories and considers environmental adaptation of species.
Taxonomic Classification

- For a start, scientists split species into three large groups after the usual beginning of all life. Bacteria, Archaea,
and Eukarya are groups called domains. After domains, the following categories of increasing specificity are kingdoms:
phylum, class, order, family, genus, and species
Phylogeny

-Phylogeny is the study of relationships and their evolutionary development among different groups of
organisms. A phylogeny is commonly represented by a phylogenetic tree called a tree diagram.
An early example of a phylogenetic tree is the "Tree of Life" by Darwin

Cladistics

-The most common way to integrate information into phylogenetic trees is called cladistics. Based on features of
ancestor and descendant species, cladistics explains theories about how organisms are linked. In
the 1950s, a scientist named Willi Hennig established cladistics. It became very popular over the
next few decades, and is still used widely today

Genetic engineering
- involves the use of molecular techniques to modify the traits of a target organism
 - Genetic engineering includes classical breeding which is considered as the traditional way of genetic
engineering which practices the mating of organisms with desirable qualities and Recombinant DNA technology
(rDNA), a modern technique of genetic engineering. Recombinant DNA technology is the joining together of DNA
molecules from two different species.

I. Cutting

- or cleavage of DNA by restriction enzymes (REs) as shown on steps 2 on the diagram. Restriction enzymes are
called ‘molecular scissors’ cutting the DNA at specific target sequences leaving a single-stranded overhang at the
site of the cleavage (step 2). These overhangs of the donor DNA (gene of interest) will be paired with other
overhangs (vector DNA).

II. Selection

- of an appropriate vector or vehicle which would propagate the recombinant DNA (shown on step 1). The most
commonly used as vectors are plasmids (circular DNA molecules that originated from bacteria, viruses and yeast cells).
Plasmids are not part of the main cellular genome, but they carry genes that provide the host cell with useful properties
such as drug resistance, mating ability, and toxins production. They are small enough to be conveniently manipulated
experimentally and furthermore, they will carry extra DNA that is spliced to them.

III. Ligation

- (join together) of the gene of interest (eg. from animal) with the vector (cut bacterial plasmid) as shown on step
3 of the above diagram. The resulting molecule is called recombinant DNA. It is recombinant in the sense that it
is composed of DNA from two different sources.

IV. Transfer

-of the recombinant plasmid into a host cell (that would carry out replication to make huge copies of the
recombined plasmid). In the above diagram as shown in steps 4, the host cell is a bacterium known also as
recombinant bacterium which will undergo cloning or replication of recombinant DNA

V. Selection

- process to screen which cells actually contain the gene of interest. The next step after cloning, therefore, is to
find and isolate that clone among other members of the library. If the library encompasses the whole genome of
an organism, then somewhere within that library will be the desired clone.

VI. Sequencing of the gene

- to find out the primary structure of the protein. Once a segment of DNA has been cloned, its nucleotide
sequence can be determined. The nucleotide sequence is the most fundamental level of knowledge of a gene or
 genome. It is the blueprint that contains the instructions for building an organism, and no understanding of
genetic function or evolution could be complete without obtaining this information.

*After the process of recombinant DNA, these plasmids or gene copies will now be introduced to its host
organisms to confer upon them the desired trait. A gene for pest resistance for example, as shown from the
image above, may be isolated, cloned and inserted into plant cell. Alternatively, bacterial cells may express the
inserted gene in order to produce protein products. Some important human proteins like hormones and
enzymes are produced by this technique.

These are some ways in which these plasmids may be introduced into host organisms;
1. Biolistic.

-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.

2. Plasmid insertion by Heat Shock Treatment.

- Heat Shock Treatment is a processused 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
CaCl2) 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 30min. 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 are taken into the cells
by this process. The cells that took up the plasmids acquire new traits and are said to be “transformed”.

3. 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.