Chapter 2.

BIOTECHNOLOGY
GEC 6 - Science, Technology, and Society
 INTRODUCTION
At home we prepare food items such as
yoghurt (curd), cake, bread, idli and
dosa etc. by the action of tiny
Modern techniques in biotechnology
microorganisms, the bacteria and fungi.
are programming microorganisms for
Brewers use yeast (a kind of fungus) to
this task. Biotechnology is applied in
make beer. Antibiotics such as penicillin
many areas to produce foods and
are obtained from certain fungi.
medicines, in the development of new
Nowadays, biological processes such as
diagnostic tools, gene therapy, DNA
fermentation by microorganisms is
finger-printing for forensic purposes
being used in industry on a commercial
etc.
scale for making food, drinks, drugs
(medicines) and industrial chemicals.
LEARNING OUTCOMES:
Discuss what is all about
biotechnology, and its application
in various areas
Trace the history of Biotechnology
through the years
Identify the advances in science
and technology have brought more
and better application of
biotechnology in various areas.
Discuss the ethics and implications
of GMOs and potential future
impacts
 WELCOME TO THE WORLD OF
BIOTECHNOLOGY
Biotechnology is many things to different people-
even if they are not aware that the product they are
using results from a biotechnology process. Our
ancestors were pioneers of biotechnology. In the
days before refrigeration and canned goods, people
looked for ways in which they could store their
surplus food. They found that when fish was stored
in jars with lots of salt and left for a while, the flesh
of the fish would become liquefied resulting into a
liquid which when added to food would make it
tastier. We call this liquid fish sauce or patis. Patis
is a popular condiment. The residue is also used as
a condiment and is called fish paste or bagoong.
It was common practice for our ancestors to
tap the flower buds or inflorescence of the
coconut tree or the nipa palm tree to collect
and store the fresh sap in a bamboo tube to
produce tuba wine. They found that tuba,
when further stored, could also turn sour to
make vinegar or suka. However, when
distilled, tuba could turn into a warming and
intoxicating drink called lambanog. Pigafetta,
the writer Magellan brought along on his trip
in the 16 century, recorded that Magellan
was gifted with a jarful of arak, or distilled
liquor when he landed in Homonhon in the
Visayas. He noted that the natives were great
drinkers, not only of arak, but also of tuba,
This arak was also very strong and could be
used to clean wounds.
Now, we know that such products were obtained with the help of very
small organisms (microorganisms) found almost everywhere-in fish, in
plants, in the soil, etc. When microorganisms break down sugars in the
coconut water or sap, alcohol is produced. Some microorganisms that
can withstand high salt condition and enzymes in the fish can break
down the proteins in the fish muscles into a tasty (and salty) liquid! These
organisms are invisible to the eye but can be seen with the help of a
microscope. The processes we have described fall under the technology
called fermentation, a kind of traditional biotechnology.
As man started to build communities, as he
became more domesticated and needed more
food to feed his family and the animals he
was taking care of, he started improving
crops-selecting for crops that gave better
looking or bigger fruits, higher yields, those
that resisted insect pests and pathogens, and
crops that provided better quality foods or
feeds. Similarly, he improved animal strains-
selecting cows and goats that gave more
milk, pigs that gave more lean meat, fowls
like chicken that gave more eggs or better
meat, and others. Improvement of crops
(plant breeding) and animals (animal
breeding) is also a form of biotechnology.
 Thus, biotechnology is defined simply as any
biology-based technology which uses organisms or
their parts to make or modify products, or improve
plants, animals and microorganisms.
 BIOTECHNOLOGY
THROUGH THE YEARS
From as far back as 4000 BC, man had learned to
prepare foods and beverage using microorganisms.
At that time, he did not know why he could make
bread rise or prepare intoxicating drinks from fruit
juices! Let us trace how biotechnology progressed
from the ancient times to the present.
BIOTECHNOLOGY
HISTORY OF
4000 BC
Egyptians discovered the use of yeasts to make their
bread rise and to prepare wine. Yeasts break down sugar
to carbon dioxide and water. The carbon dioxide makes
bread rise and wine and beverages bubbly.

4000
3000 BC
BC
Egyptians discovered
Peruvians started the usethe
to improve of yeasts
potato.toThey
make theirit
made
bread rise and
larger and ableto
toprepare wine.
resist frost. Yeasts
They usedbreak down sugar
different
to carbon dioxide
techniques and water. The
and crossbreeding carbon dioxide
in improving makes
the potato.
bread rise and thus
The Peruvians winecontributed
and beverages bubbly. classic
to developing
biotechnology techniques.

1763 BC
Sumerians invented the process of brewing beer.
BIOTECHNOLOGY
HISTORY OF
1683
Antonie van Leeuwenhoek invented the microscope. This
invention allowed scientists to study tiny objects, micro-
organisms and even molecules that our naked eyes
cannot see.

1861
4000 BC
Louis Pasteur invented a process that destroys harmful
Egyptians
bacteria indiscovered the use
milk and other of yeasts
products but to make
does nottheir
affect
bread riseofand
the taste theto prepare
food or itswine. Yeasts
nutritional breakThis
value. down sugar
process
to called
is carbonpasteurization.
dioxide and water. The carbon dioxide makes
bread rise and wine and beverages bubbly.
1865
Gregor Mendel was a monk and scientist who studied
easily measured color, shape and length. He discovered
the basic laws of genetics but this was largely
unacknowledged until the following century when such
laws were re-discovered by other scientist.
BIOTECHNOLOGY
HISTORY OF 1879
Williom James Beal developed the world's first hybrid
corn by crossing white and yellow corn plants which
produced yellowish white

4000 BC
1944
Avery, Macleod
Egyptians discovered
and McCarty
the use established
of yeasts to DNA
makeas
their
the
bread rise and
hereditary material
to prepare
or the gene
wine. Yeasts break down sugar
to carbon dioxide and water. The carbon dioxide makes
bread
1865rise and wine and beverages bubbly.
Barbara McClintock discovered "jumping genes" in her
studies of maize. Earlier, scientists thought that the gene
would always stay in one place but McClintock
discovered that jumping genes naturally occur.
BIOTECHNOLOGY
HISTORY OF
1953
James Watson and Francis Crick discovered the double
helical nature of DNA, The double helix structure explains
how the DNA could replicate or produce identical copies of
itself, and how the genetic information in the gene is
expressed. This discovery opened up man avenues of study
and affected almost all aspects of the sciences.
4000 BC
1973
Egyptians discovered the use of yeasts to make their
bread
Stanleyrise and and
Cohen to prepare
Herbertwine.
BoyerYeasts break down
demonstrated that asugar
gene
to
onecarbon dioxide
organism can beand water. The
introduced carbon
into dioxide
another makes
organism and
bread rise andThey
be expressed. winecut
and beverages
the bubbly.
gene for the ribosomal RNA from
toad DNA with a restriction enzyme, inserted the gene in a
small circular extra chromosomal DNA called plasmid, and
joined them together using another enzyme called ligase.
The plasmid was introduced into bacteria E. coli where it
was multiplied and the toad gene was expressed. This is the
start of recombinant DNA technology or genetic
engineering.
BIOTECHNOLOGY
HISTORY OF
1982
Human insulin produced by bacteria as a result of genetic
engineering became the first product of modern
biotechnology to be commercially released.

1987
4000 BC
A crime suspect in the UK was convicted on the evidence
of genetic discovered
Egyptians fingerprinting.
the Individuals have
use of yeasts specific
to make DNA
their
pattern which
bread rise and can differentiate
to prepare wine. them from
Yeasts others,
break down sugar
although
to carbonthey will and
dioxide share common
water. The patterns with close
carbon dioxide makes
relatives.
bread rise and wine and beverages bubbly.

1990
Recombinant chymosin produced in E. coli was released
commercially. This enzyme is used to coagulate milk to
produce cheese.
BIOTECHNOLOGY
HISTORY OF
1991
The first transgenic animal, Tracey, a sheep, was born.
Tracey had human genes which enabled her to produce
human therapeutic proteins in her milk

1994
4000 BC
The FLAVRSAVR" tomato with delayed ripening trait and
higher solids
Egyptians was the first
discovered genetically
the use of yeastsmodified
to makecrop
theirto be
commercially
bread rise andreleased.
to prepare wine. Yeasts break down sugar
to carbon dioxide and water. The carbon dioxide makes
1996
bread rise and wine and beverages bubbly.
Several genetically modified or biotech crops were
introduced into the market- herbicide tolerant (HT)
soybean, corn, rapeseed and cotton, and insect-
protected or BT corn and cotton.
BIOTECHNOLOGY
HISTORY OF
1997
Dolly the sheep, the first cloned animal, was born
developed by Dr. Ian Wilmut and his colleagues at Roslin
Institute near Edinburgh, Dolly was cloned from an udder
cell of an adult sheep; the udder cell was fused with an
egg cell whase nucleus had been removed. The fused
udder
4000cell-egg
BC was allowed to divide and then implanted
into a different sheep.
Egyptians discovered the use of yeasts to make their
bread
2002 rise and to prepare wine. Yeasts break down sugar
to carbon dioxide and water. The carbon dioxide makes
The
breadhuman genome
rise and wine was
and successfully sequenced.
beverages bubbly.

2003
The area planted to genetically modified crops (herbicide
tolerant soybean, Bt corn and Bt cotton, rapeseed)
increased to 81 million ha in 17 countries in the world
BIOTECHNOLOGY
HISTORY OF
2006
The top ten biotech health products for anemia, diabetes,
cancer, arthritis, etc, had a market worth US$33 B. The
first biotech drug, an anti-clotting protein, produced in
the milk of goats was approved for release in 2006 in
Europe and in 2009 in the USA.

4000 BC
2009
the first blue
Egyptians discovered
rose wasthe
released
use ofcommercially
yeasts to make
in their
Japan
breadalmost
after rise and
twotodecades
prepare of
wine.
research.
Yeasts break down sugar
to carbon dioxide and water. The carbon dioxide makes
2012
bread rise and wine and beverages bubbly.
Ten years after the completion of the human genome, its
contributions to biomedical research continue to
increase, allowing us to better understand the biological
functions encoded in the genome, to learn more about
the basis of diseases and cancer and the evolution and
history of man
ADVANCING THE
FRONTIERS OF
BIOTECHNOLOGY
Advances in science and technology have brought
more and better applications of biotechnology in
health and medicine, food and agriculture,
environment and industry. Biotechnological
comprise two types conventional and modern.
Both cover a wide range of applications
techniques and processes application of
biological processes, but the main difference is
that modern biotechnology covers those
techniques and processes which focus at
molecular- or gene level. These include DNA
marker technology, genomics and recombinant
DNA technology or genetic engineering.
 VACCINES AND ANTIBODIES
Remember when you were a baby, your mother brought you to
the doctor for vaccination against diseases like measles,
polio, tuberculosis, etc. Vaccines are made of heat-treated
(dead) pathogens or mild strains of such pathogens that
cause the disease. When injected into your body, a vaccine
will stimulate the production of antibodies by your own body
to protect you against a specific disease. Now, modern
vaccines are made of only parts of the organisms like the cell
wall, or even specific proteins on the cell wall which can
stimulate production of specific antibodies. Antibodies are
the globulin proteins in the blood serum of animals and are
called immunoglobulins. They protect man against invading
pathogens that cause diseases.
 PLANT BIOTECHNOLOGY
For thousands of years, man has developed various ways of
propagating and multiplying plants. He found that some
plants are easily propagated by planting their seeds, Plants
coming from seeds may be similar or even different from the
source or mother plant. This kind of propagation is called
RECAP 01 RECAP 02
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 PLANT BIOTECHNOLOGY
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Summaries and recaps also reinforce the
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 ANIMAL BIOTECHNOLOGY
Advanced techniques to improve animal strains have been
available for me decades. One of these is embryo transfer
(ET), which is a widely utilized technology for large
animals such as cattle, buffaloes or carabaos and gots.
The Biotechnology Research Team of the Philippine
Carabao Center based in Nueva Ecija, has successfully
produced healthy elite breeds of water buffaloes
(carabao from in vitro-produced- frozen-thawed
embryos.
The in vitro embryo production was done in Aurangabad,
India and the frozen embryos transported to the country.
They were thawed and implanted in surrogate female
native carabaos.
In 2004, six healthy calves including Glory were born.
These are now used as breeding animals to improve the
local buffaloes in their ability to produce milk. The water
buffaloes from India belong to the Indian Murrah breed
and are excellent milk producers.
 ANIMAL BIOTECHNOLOGY
Cloning of animal twinning (or production of twins).
Under certain conditions, the unfertilized eggs of some
animals (lizards, frogs, some fishes, etc.) can grow up to
adulthood. In 1997, the first animal was successfully
cloned by Ian Wilmut and his colleagues at the Roslin
Institute in Edinburgh, U.K. through a process called
somatic cell nuclear transfer . They fused the mammary
gland cell of a Finn Dorsett sheep (a white faced-sheep)
with the enucleated egg (an egg whose nucleus has been
removed) of a Scottish blackfaced ewe by using
electrical pulses.
ANIMAL
BIOTECHNOLOGY
The famous Dolly the sheep was
born using this cloning technology
but only after 276 attempts Dolly
was shown to be genetically
identical to the Finn Dorsett sheep
and not to the blackfaced ewe.
Animal cells can be cultured from
tissues of specific organs like
skin, liver, etc. Some types of
mammalian cells have been
utilized as bioreactors which
produce various health products
such as antibodies, and antiviral
and anticancer interferon through
the technique of recombinant DNA
technology or genetic
engineering.
RECOMBINANT DNA TECHNOLOGY
OR GENETIC ENGINEERING
In the early 1970s, laboratory techniques that allowed
the cutting of DNA at specific sites, joining the DNA
pieces together, and introducing the rejoined DNA in
bacteria to be multiplied, became available. Thus,
recombinant DNA technology or genetic engineering
was launched! This new technology has brought about
the production of human health products like insulin,
hepatitis vaccine, and a cancer-fighting substance
called interferon in bacterial and mammalian cells The
new technologies also brought about the ultimate
diagnostic tool to identify human individuals through
DNA fingerprinting. Genetic engineering has also
introduced improved traits in crops that help the farmer
and the consumer. This technology will also enable the
use of animals and crops as factories to produce
important health and industrial products.
HUMAN CLONING AND STEM CELLS
FOR THERAPY
When the news of the cloning of Dolly the sheep came out in
1997, many believed that within the decade, the more shocking
announcement of the cloning of a human being would occur. In
2002 and 2003, there were claims of pregnancy initiated through
human reproductive cloning and births of the first human clones.
However, these claims have not been scientifically substantiated
and have been treated with much skepticism. Most people and
many countries agree that human cloning to produce humans (or
human reproductive cloning) is unethical and should not be done
for any purpose. The United Nations General Assembly adopted
on August 3, 2005, the "United Nations Declaration on Human
Cloning, by which Member States were called on to adopt all
measures necessary to prohibit all forms of human cloning
inasmuch as they are incompatible with human dignity and the
protection of human life.
HUMAN CLONING AND STEM CELLS
FOR THERAPY
However, the other type of human cloning called human
therapeutic cloning promises to have a wide range of useful
applications in medicine. With human therapeutic cloning, DNA
from a sick person can be used to grow an embryonic clone
obtained by somatic cell nuclear transfer method. The extracted
DNA from a sick person is inserted into an enucleated donor egg.
The resulting egg then divides like a fertilized egg and forms an
embryo. But instead of inserting the embryo into a surrogate
mother, cells from the embryo are used to grow stem cells in
vitro.
HUMAN CLONING AND STEM CELLS
FOR THERAPY
Stem cells are unprogrammed cells in the human body which
have the ability To change into 220 types of cells. Stem cells are
thus described as pluripotent. These stem cells con therefore be
used as a human repair kit. They have the potential to become
bone, muscle, cartilage and other special types of cells.
Replacements for organs such as heart, liver and skin can be
obtained from ste cells. These stem cells can also be
transplanted back to the donor to treat diseases which at this
time do not have cures such as Alzheimer's, Parkinson's, heart
failures, spinal cord injury, etc.
HUMAN CLONING AND STEM CELLS
FOR THERAPY
Another important potential use of stem cells is for testing new
medications for safety and effectiveness. For example, an anti-
tumor drug can be tested on a cancer stem cell line to test if the
drug can stop the cancer cells from growing.

To get stem cells, scientists can use embryos obtained

fertilization or embryos cloned by somatic cell nuclear transfer as
described above. Either way, scientists, as they move forward to
understanding and manipulating stem cells, inevitably destroy
the embryo from which the stem cells are derived. This is a highly
controversial ethical issue because of the question of when life
starts.
HUMAN CLONING AND STEM CELLS
FOR THERAPY
Because of the ethical issues surrounding the use of stem cells
from embryos, scientists are exploring less controversial
materials to study stem cells. They are using adult stem cells that
can be derived from umbilical cord, peripheral blood, or bone
marrow. These adult stem cells can be trained to act like
embryonic stem cells and be able to differentiate to specific cell
types under defined conditions. However, it will still take
decades before the benefits of stem cell therapies can be
enjoyed by the general public.
 BASIS OF GENETIC ENGINEERING
To better understand the process of genetic engineering or recombinant
DNA (deoxyribonucleic acid) technology, let us review some basic
biological principles.

Cells of different organisms like microorganisms, plants and animals are

similar.

Animal cells, plant cells and bacterial cells contain the same biomolecules
like proteins, DNA, carbohydrates, lipids etc. They have similar organelles
or bodies which have similar functions. However, the bacterial cell does
not have a distinct body that contains its DNA, unlike animal and plant
cells which have the nucleus which contains their DNA. They follow the
same physical, chemical and genetic laws. Thus, if a gene is transferred
from one organism to another, and is integrated into the genetic material
of that organism, it may work and has been proven to work!
 BIOLOGICAL MOLECULES
1. Proteins are large 2. Nucleic acids can be
molecules made up of one deoxyribonucleic acids
or more chains of amino (DNA) or ribonucleic acids
acids. They perform a (RNA). DNA is the chemical
wide variety of activities which carries the
in the cell. Enzymes are instructions for all the traits
protein catalysts of of an organism. DNA occurs
biochemical reactions, for as a double-stranded helix
example, proteases digest or a double helix. RNA or
the food we eat. Muscles ribonucleic acid is a single
that help us move are stranded chemical similar to
made up of proteins. Many DNA. RNA serves as a
hormones such as messenger of the DNA's
somatostatin, insulin, genetic message to the
growth factors, etc are cytoplasm of a cell where
proteins. Our hair is made proteins are synthesized.
up of proteins.
 BIOLOGICAL MOLECULES
3. Carbohydrates are
represented by the sugars
and their long chains. The
most abundant sugar or
monosaccharide is glucose
which is the major fuel or
source of energy of the
organism. The long linear or
branched chains of
polysaccharides serve as
chief fuel storage form and
as structural elements.
4. Lipids are water-insoluble
organic biomolecules whose
important biological
functions are (1)as structural
components of membranes,
(2) forms of metabolic fuel
for storage and transport
(3)as coating to protect the
surface of many organisms
and (4) as components of
cell surface involved in
recognition of cell,
specificity of species and
immunity of tissue.
 WHAT IS DNA?
The DNA is like a string of notes in specific
order. This string notes becomes melody
when expressed vocally by a singer, or on a
piano by a pianist, or by any other musical
instrument.

A DNA consists of two strands (string), each

string having chemical components called
bases joined together by a sugar phosphate
backbone. The two strands run in opposite
direction. The double strands are held
together by hydrogen bonds linking specific
bases. Adenine (A) pairs with Thymine (T).
Cytosine (C) DNA strand in the double helix
are pairs with Guanine (G).
 WHAT IS DNA?
The pairing of the bases is specific and complementary. Thus,
one strand of a DNA is said to be complementary to its partner
strand in the double helix and both strands will actually bear the
same genetic information. The double helix structure of the DNA
which was discovered by James Watson and Francis Crick in
1953 explains why in the process of DNA replication, the
resulting daughter DNA molecules are identical to the parental
DNA helix.

The letters A, T, G and C form the code of life. In the human

genome, there are about 2.9 billion base pairs which are wound
into 24 bundles called chromosomes. About 22,000 genes are
written in the human DNA which code for the many proteins that
help build and maintain our bodies
WHAT IS DNA?

The figure shows the

location of the DNA in
the nucleus of a typical
cell. Unicellular cells do
not have a distinct
nuclear body. The
double helical DNA in a
cell when stretched
measures about two
meters. Genes are
portions of the DNA
which carry information
that can be carried from
gene from generation to
generation.
 THE INFORMATION FOR A
TRAIT IS ENCODED IN THE
DNA CALLED THE GENE
The information for a trait (like the color or the pigment
of your eyes) is encoded in the molecule called DNA or
deoxyribonucleic acid and is called a gene. In a cell, the
information for a gene, is expressed into a protein
through o messenger molecule called messenger
ribonucleic acid or mRNA, We call protein the primary
gene product. Thus, genes contain instruction for the
production of proteins that are involved in different types
of activities that make our bodies function.
 THE INFORMATION FOR A TRAIT IS
ENCODED IN THE DNA CALLED THE GENE
We also know that proteins have several types of
functions like providing structure (muscles), facilitating
or catalyzing chemical reactions (as enzymes),
transporting materials and in communicating with other
cells (hormones). You are familiar with hemoglobin, the
protein carrier of oxygen in the blood. Specific enzymes
produce the pigment that gives the color of our eyes or
the keratin of our hair and nails. Enzymes catalyze
specific reactions resulting into products which
contribute to the creation of a trait. While a simple trait
may be encoded in a single gene, several genes
contribute to and affect complex traits such as human
intelligence and crop yield.
 THE INFORMATION FOR A TRAIT IS
ENCODED IN THE DNA CALLED THE GENE
The information in the gene is transferred or transcribed
to a complementary single strand called messenger RNA
or mRNA, base U (for Uracil) is used instead of T. This
process is called transcription.

The mRNA travels from the nucleus to the cytoplasm of

the cell and is translated to protein. This process is
called translation.
 GENETIC ENGINEERING OF
MICROORGANISMS, PLANTS
AND ANIMALS
Recombinant DNA technology or genetic engineering is a
method that allows the combination of genes in a fest
tube to form a hybrid DNA. It allows the transfer of a
specific gene (from the same or different organism) to
produce a new trait in an organism.

The basic steps involved in genetic engineering are the

following: (1) extraction of the DNA from the source
organism, isolating and cloning the gene from the DNA:
(2) design and preparation of the "gene construct" or
"hybrid DNA“ or "recombinant DNA", (3) delivery of the
gene construct to the new host organism. and (4)
checking the presence and expression of gene in new
host organism. In the 1970s, the tools to make
recombinant DNA technology happen were already
available and hastened its development
To design the gene construct, one considers the different
parts of a gene the promoter region, the so-called structural
gene part and the terminator sequence (Figure 8). As the
term connotes, the promoter region contains elements or
sequences which drive the reading of the gene, instructing
when and where the gene will be expressed. For example,
the promoter can direct the expression of the gene in all
parts of the organism or in specific tissues or organs (seeds,
leaves, etc.), at all times or only at night or daytime or at a
specific stage of the development of the organism. The
structural gene part provides the information for the trait
itself (e.g., a specific protein) and the terminator sequence
marks the end of the transcription of the gene.
 TOOLS OF RECOMBINANT DNA
TECHNOLOGY
1. Restriction enzymes or restriction endonucleases
cut DNA at or near specific nucleotide sequences.
2. DNA ligase joins fragments of DNA.
3. DNA polymerase catalyzes the synthesis of DNA,
RNA polymerase catalyzes the synthesis of VNH
4. Electrophoresis separates charged molecules based
on charge (neutral, positive and negative) and size.
Charged molecules move on a medium to the
opposite pole when current is applied.
5. Polymerase chain reaction is a technology which
allows the amplification of parts of DNA by in vitro
replication that involves repeated denaturation,
primer attachment and synthesis of DNA.
MICROOORGANISMS
ENGINEERING OF
GENETIC
The steps involved in the process of genetic
engineering of microorganism are shown in the figure
below to illustrate the introduction of the human insulin
gene into bacteria.

4000 BC
Egyptians discovered the use of yeasts to make their
bread rise and to prepare wine. Yeasts break down sugar
to carbon dioxide and water. The carbon dioxide makes
bread rise and wine and beverages bubbly.
MICROOORGANISMS
ENGINEERING OF
GENETIC
STEP 1
The gene of interest, in This case, human insulin, is
isolated from the human DNA and cloned. [Cloning of
DNA or a gene means making multiple exact copies of
the DNA or gene]
4000 BC
STEP 2 discovered the use of yeasts to make their
Egyptians
bread rise and to prepare wine. Yeasts break down sugar
The human insulin gene is inserted into a vector-a
to carbon dioxide and water. The carbon dioxide makes
plasmid- at specific sites cut by enzymes called
bread rise and wine and beverages bubbly.
restriction enzyme, A plasmid is a genetic element
(DNA) which can carry a DNA fragment into a recipient
cell like a bacterium. The plasmid contains the
promoter elements and terminator sequence that will
work for the host organism, in this case, bacterial cells.
MICROOORGANISMS
ENGINEERING OF
GENETIC
STEP 3
The recombinant plasmid is introduced into bacterial
cells by physical means. The bacterial cells are
subjected to heat and then put on ice. This will result
into formation of pores in the cellular membrane that
allow the DNA to get inside the cell. This process is
called transfection or transformation. The recombinant
4000
plasmid BCis not incorporated in the chromosome of the
bacteria
Egyptiansbut it is multiplied
discovered the useinto many copies
of yeasts to makeintheir
the cell.
bread rise and to prepare wine. Yeasts break down sugar
STEP
to carbon4 dioxide and water. The carbon dioxide makes
bread rise and
This results wineproduction
in the and beverages bubbly.
of many copies of the
inserted gene (the human insulin gene) and the
production of human insulin by the bacterial cells. This
is called cloning of the insulin gene and this is how
human insulin is produced by bacterial cells. The same
process is basically followed for the cloning of other
genes in microorganisms to produce various products.
 STEP 1
The gene or genes needed are isolated and cloned.

GENETIC STEP 2
ENGINEERING The gene is inserted in an appropriate vector to form
the gene construct. A vector is like a vehicle or carrier
OF PLANTS and has the necessary regulatory elements such as the
promoter and terminator which con make the gene
work. The promoter will tell when and where the gene
will be expressed and how many copies of the protein
will be produced. The terminator will command the end
of expression or reading of the gene. A selection gene
marker is also usually in the gene construct. This will
help in determining which of the bombarded tissues
have incorporated the gene construct in their DNA.
 STEP 3
The gene construct is delivered to the plant cell by
either of two methods. One is by coating the gene or
GENETIC DNA on tungsten or gold particles and delivering the
DNA-coated particles into plant tissues by using a
ENGINEERING particle bombardment device. This device is attached
to a pressurized tank containing helium gas which
OF PLANTS forces the DNA-coated particles into the tissues with
pressure. The other method involves inserting the gene
construct into Agrobacterium tumefaciens which is then
used to infect a plant and eventually transfer the gene
construct and its other genes to the plant genome. The
Agrobacterium is commonly found in nature. If you find
galls or swellings on plants, most probably, this is due
to the infection of the plant with Agrobacterium.
 STEP 4
The next step is to determine which among the plant
GENETIC cells have integrated the introduced gene, The
selection marker gene will do this! For example, if the
ENGINEERING selection marker gene is an antibiotic resistance gene
marker, cells which have this gene will be able to
OF PLANTS survive in a medium containing such antibiotic. These
cells are termed transformed or transgenic, [Another
type of selection marker gene is the green fluorescent
protein or GFP marker; cells that integrate this in their
DNA will produce this protein which gives off green
fluorescence when beamed under a UV light]
 NOTE THAT:
The transformed or transgenic plant tissues
are allowed to grow and regenerate to
complete plants. The breeder will now
GENETIC screen the resulting plants for the desired
ENGINEERING trait and evaluate as well their agronomic or
horticultural traits. The breeder will select
OF PLANTS lines which have the desired traits and are
stable. The molecular biologist/biochemist
will determine the presence of the inserted
gene and other biochemical characteristics
of transgenic plants. These steps as are
basically the same for all genetically
modified, or transformed or transgenic
plants.
GENETIC Here, we are introduced to different terms to describe
plants, animals or microorganisms or products that have

ENGINEERING resulted from a genetic engineering process genetically

modified, genetically engineered, transformed and
OF PLANTS modern biotech plants or products. bioengineered,
transgenic, transformed and modern plants or products
GENETIC ENGINEERING
OF ANIMALS
With the advances in science and technology, it is now possible
to produce transgenic animals which can provide economical
means of producing life-saving health products. As in the case of
recombinant microorganism or transgenic plant, transgenic
animal has a new gene introduced in its genome by in vitro
methods.

The basic requirements for genetic engineering of animals are

similar to those for genetic engineering of microorganisms and
plants. We need to: (1) identify, isolate and clone the gene of
interest: (b) design and prepare the gene construct or
recombinant DNA; (c) deliver the gene construct to the animal
system and (d) check the presence and expression of the gene in
the new host. The transferred gene should be stably integrated in
the genome of the host animal and be inherited by its progenies.
GENETIC ENGINEERING
OF ANIMALS
How are transgenic animals produced?
The figure shows the different steps involved in genetic engineering
of animals using the microinjection method to deliver the gene,
After the scientist has identified and isolated the gene to be
introduced to the animal for the desired trait, this gene or DNA is
injected into a fertilized egg of the animal using a very fine glass
needle viewed under a powerful microscope. The gene is
incorporated into the DNA of the animal's genome and encodes the
instruction for the new trait. If the gene is integrated in the
fertilized egg before it divides, then every cell in the animal will
contain the gene, and the gene will be passed on by the animal to
its offspring. When the fertilized egg or zygote has divided into
several cells, it is implanted into a surrogate mother, where it
develops to full term and is delivered. This method has been used
to develop transgenic animals such as cattle, sheep and goat which
can produce health products in the animal's milk.
GENETIC ENGINEERING
OF ANIMALS
How are transgenic animals produced?
So far, the only commercially available transgenic animals are
transgenic mice and the GloFish. Transgenic mice are used as
experimental animals to study The effects of overexpressing or
misexpressing foreign or native genes at specific Stages or
locations in the animal. GloFish is an ornamental aquarium fish
called zebra fish which gives off attractive fluorescing or glowing
colors of red, blue, black, green and yellow (Figure 13). This was
developed by microinjection of one- or two-cell embryos of the
zebra fish with a gene construct containing the gene for green
fluorescent protein (GFP), red fluorescent protein (RFP) and yellow
fluorescent protein (YFP) and the promoter of the zebrafish muscle
protein myosin
GENETIC ENGINEERING
OF ANIMALS
How are transgenic animals produced?
Because of the fluorescing nature of the
introduced proteins(GFP, RFP, and
YFP), the transgenic lines could be
screened by observing the fluorescing
proteins in the offsprings. More
recently, teo other types of glofish were
released commercially, these are
GloFish Tetra and the GloFish Electic
Green Barb. The GloFish is available
commercially in many states in the
United States of America.
Some of the
transgenic fish which
are at various stages
of development are
fish such as tilapia,
flounder and salmon
which grow faster than
nontransgenic fish,
and fish that are more
tolerant to cold or
resistant to certain
diseases.
The first recombinant drug produced by an
animal, a transgenic goat, is ATryn®
Antithrombin. ATryn is indicated to prevent
perioperative and peri-partum abnormal
blood clotting events in patients who have
hereditary anti-thrombin deficiency. To
produce this product, the transgene
construct contains the human gene for
antithrombin, the promoter of the milk
protein casein, and other elements to
ensure high production in milk of 2 g per
liter to as high as 20 g per liter. This product
was approved for marketing in Europe by
the European Medicine Agency in 2006 and
in the US by the US Food and Drug
Administration in 2009.
 BIOTECHNOLOGY IN THE
PHILIPPINES: WAY TO GO
Even before 1979, different
biotechnology researches were
already being conducted at the
University of the Philippines Los
Baños (UPLB). The year 1979
witnessed the establishment of the
National Institute of Biotechnology
and Applied Microbiology (BIOTECH)
at UPLB by a Presidential Decree
(PD) under President under
Ferdinand E. Marcos.
 BIOTECHNOLOGY IN THE
PHILIPPINES: WAY TO GO
From 1986-1992, biotechnology
was a flagship program of the S&T
program under President Corazon
C. Aquino. The Department of
Science and Technology had
identified biotechnology as a
strategic tool for achieving
sustained economic development."
The National Committee on
Biosafety of the Philippines (NCBP)
was established in 1990 by virtue
of Executive Order (EO) 430
Biotechnology remained as a major
S&T program from 1992-1998
President Fidel V. Ramos. During this
time, a network of Biotech institutes
was established at the University of
the Philippines campuses by then UP
President Emil Q Javier. Dr. EQ Javier
was also instrumental in the
establishment of the first BIOTECH
Institute at UPLB in 1979. In 1992, the
Institute of Plant Breeding of the
College of Agriculture, UPLB, was
mandated by the Seed Industry
Development Act of 1992 to lead in
plant biotechnology activities.
 BIOTECHNOLOGY IN THE
PHILIPPINES: WAY TO GO
In 1997, the Agricultural and Fisheries Modernization Act of 1997
(AFMA) was enacted into law. The AFMA recognized the potentials of
biotechnology to contribute to the modernization of agriculture and
fisheries and thus included a provision for biotechnology.

During the term of President Joseph E. Estrada, biotechnology was

institutionalized in government programs in 2000.

In July 2001, President Gloria M. Arroyo issued a policy statement

on modern b biotechnology: "We shall promote the safe and
responsible use of modern biotechnology and its products as one of
several means to achieve and sustain food security, equitable access
to health services, sustainable and safe environment, and industry
development."
 BIOTECHNOLOGY IN THE
PHILIPPINES: WAY TO GO
In 2002, Administrative Order No. 8 of the Department of Agriculture
was issued by DA Secretary Leonardo Q Montemayor. This AO covers
the rules and regulations for the importation and release into the
environment of plants and plant products derived from the use of
modern biotechnology.

On November 5, 2013, in a speech delivered during an international

rice conference, President Benigno Aquino III acknowledged the role
of biotechnological research in developing more cost effective
varieties of rice like Golden Rice and Submarino Rice high quality
corn we produce." and the role of scientists "to build a more
sustainable future for all our peoples.“
The following provides a brief overview of the
R & D and commercial aspects of various
biotechnologies in different areas in the
Philippines, Table below lists biotechnologies
utilized in the country and indicates imported
and these developed and produced locally.
HUMAN AND AND
Existing products
ANIMAL HEALTH

Only a few animal vaccines (against hemorrhagic septicemia,

pasteureloses, ND, fowl pox, and anthrax) are locally manufactured by
Vaccines
conventional methods. Most of animal and human health vaccines are
imported.

Mostly imported; locally developed diagnostics for mycotoxins and red

tide toxins await commercialization; PCR-based diagnostics for
Diagnostic
Salmonella developed by UPLB BIOTECH; PCRbased kit for dengue
developed by UP Manila BIOTECH awaiting commercialization

Bioactive Therapeutic All imported: Insulin, alpha interferon, hepatitis B surface antigen-
Protein based vaccine, erythropoietin, filgrastim, others

Blood and blood products, monoclonals, bioactive peptides, effective

Other product
biotech drug delivery systems are all imported.
AGRICULTURAL AND LIFE
Existing products
SCIENCE

Wine (tapuy, tuba, basi), cheese, bread, soy sauce, vinegar,

1. Food products produced by
fish sauce (patis), fish paste (bagoong), achara, various "buro“
fermentation / bioconversion
products, nata de coco, others

Have been developed using conventional breeding by various

2. Improved crop varieties
government research institutions and private companies.

3. Tissue culture-raised planting Tissue- cultured orchids, banana and pineapple; makapuno
materials and cut flower coconut; sugarcane

4. Biopesticides, biocontrol
BIOCON for control of nematode pests; preparations of Bt
agents

Rhizobium, etc; Mycorrhiza, Azospirillum, Bio-N,

5. Biofertilizers
Nitroplus,Cocogroe, Cocorich
AGRICULTURAL AND
Existing products
LIFE SCIENCE

6. Diagnostic Immuno-based and DNA PCR- based diagnostics for various diseases

7. Animal improvement Artificial insemination; multiple ovulation and embryo transfer

8. Animal production Sex-reversal technology; biotech-based nutrition

For varietal identification, genetic diversity analysis, markerassisted

9. Molecular markers selection or breeding, DNA barcoding for species identification,
differentiation and genetic analysis

Imported GM crops: Bt corn, herbicide-tolerant corn, GM corn with

10. GM crops both Bt and herbicide-tolerance traits, Homegrown: long shelf-life
papaya, Bt eggplant, Golden Rice
 INDUSTRIAL AND
ENVIRONMENTAL Existing products
BIOTECHNOLOGY

Mostly from sugarcane and molasses; other potential feedstocks are

1. Alcohol production
cassava, sorghum.

Technologies developed for production of a amylases,

2. Industrial enzymes proteases,cellulases, glucoamylase, lipase, pectinase, xylanase,
ligninase; most industrial enzymes are imported.

3. Amino acid production Amino acids mastly imported

4. Recovery of valueadded B-monoglycerides biopolymers such as nata de coco, applications of

products from water high strength, biodegradable membrane

Chemical fertilizer substation, composts, organic ferlitizer

5. Bioremediation
Formulation; soil amendments or supplements
PINOY GENETICALLY
MODIFIED PRODUCTS
 LONG SHELF-LIFE PAPAYA
At the Institute of Plant Breeding (IPB) at the University of the Philippines
Los Baños, researchers have developed papaya with longer shelf life using
the tool of genetic engineering to address the problem of postharvest
losses. The strategy used was to suppress the production of the hormone
ethylene involved in ripening. This was done by introducing the gene for
the ripening-related key enzyme ACC synthase into papaya in reverse or
antisense orientation (thus, the term antisense technology) which could
result in the non- or lower expression of the ripening-related ACC
synthase and therefore will also result in the lowering of ethylene
production. This project was funded by the Philippine Department of
Science and Technology-PCAARRD, The Australian Centre for
International Agricultural Research (ACIAR), the US Agency for
International Development and the Department of Agriculture
Biotechnology Program.
LONG SHELF-LIFE PAPAYA

IPB researchers representing tour different events with

good horticultural traits including delayed ripening
trait. Resistance to papaya ringspot virus (PRSV) in
papaya has also been introgressed into the transgenic
papaya resulting into hybrid papaya with both long
shelf life and PRSV resistance traits. The transgenic
papa with long shelf life and PRSV resistance is
undergoing field trails under a biosafety regime.
GOLDEN RICE WITH HIGH
PROVITAMIN A CONTENT

It is estimated that more than 125 million

children suffer from Vitamin A deficiency
(VAD) worldwide. VAD causes blindness and
premature death among children.

At the Philippine Rice Research Institute,

researchers have crossed transgenic rice
containing the genes to synthesize beta
carotene or provitamin A in the grain with local
elite rice varieties to produce local Golden
Rice lines. The Goiden Rice lines are being
field-fested in different sites in the country.
 GOLDEN RICE WITH HIGH
PROVITAMIN A CONTENT
The International Rice Research Institute is coordinating an
international network among national rice research institutes to
develop outstanding local lines of the Golden Rice.
Consumption of Golden Rice in normal amounts along with
other Foods will enable consumers to meet the daily
requirements for Vitamin A. Golden Rice was developed by
Professors Peter Beyer and Ingo Potrykus. Scientists have
succeeded in increasing the level of provitamin A by 20-fold
since the first Golden Rice was developed in 1999. The Golden
Rice containing high provitamin A will be an excellent health
food for people, in general, and for those who are suffering
from Vitamin A deficiency, in particular.
 BT EGGPLANT WITH RESISTANCE
AGAINST PEST BORERS
Eggplant is a very popular vegetable in the country planted to more than 31,000 ha.
However, eggplants are very susceptible to insect pests, primarily the fruit and
shoot borers. Damage due to borer infestation can go up to 100%: even less severe
infestation Results in fruits with many holes which cannot be sold or are sold at low
prices. Moreover, farmers resort to heavy use of chemical pesticides (40 to 80
applications) to control the borers.
 BT EGGPLANT WITH RESISTANCE
AGAINST PEST BORERS
The Bt eggplant technology provides an excellent alternative to the
ordinary plant which is heavily sprayed with pesticide. Through genetic
engineering. The gene for protein toxic only to the pest borers was
incorporated in the genome of eggplant by researchers at Mahyco in
India. The Bt eggplant produces this protein in the fruit, shoot and leaf of
the plant which when eaten by the borer will cause its death. This
technology is similar to that used in the Bt corn which is grown and
commercialized in the Philippines. The B1 eggplant technology was made
available to The University of the Philippines Los Baños through a royalty-
free license agreement. The Bt eggplant promises a good and assured
income To The farmers and will provide good quality fruits to the
consumers (Fig. 16c). Moreover, this Technology will reduce
tremendously the application of chemical pesticides which will benefit
the health of the farmers and the environment.
ACADEMIC OFFERINGS TO
PREPARE FOR CAREERS IN
MOLECULAR BIOLOGY AND
BIOTECHNOLOGY
 In the early 1980s, universities and colleges in the United
States of America started developing and offering courses
to address the needs of biotech-based industries. There
are now BS Biotechnology programs and advanced degree
programs (MS and PhD) in the US as well as in Australia.
More common is specialization in molecular biology
offered in various Universities. In the Philippines, the
University of the Philippines (UP) Diliman started offering
the BS, MS and PhD MBB (Molecular Biology and
Biotechnology) program in 1987. UP Los Baños (LB) opened
its MS MBB program in 1999 and its PhD MBB in 2002. In
October 2009, the UPLB College of Agriculture 's BS
Agricultural Biotechnology (ABT) Program was approved by
the UP Board of Regents. The BSABT curricular program
welcomed its first batch in 2010 and now has more than
250 students. Mapua Institute of Technology has been
offering a BS Biological Engineering degree since 2003.
The Visayas State University has offered BS Biotechnology
since 2007.
THANK YOU
FOR
LISTENING!