MODULE 1: BASIC BIOTECHNOLOGY

INTRODUCTION TO BIOTECHNOLOGY
A. DEFINITION OF BIOTECHNOLOGY

BIOTECHNOLOGY
 The utilization of biological processes, organisms or systems to produce products that are
anticipated to improve human lives is termed biotechnology.
 This can be defined as the engineering of organisms for the purpose of human usage.
 It can also be defined as the skill set required for the utilization of living systems or the
influencing of natural processes so as to produce products, systems or environments to
help human development.
 In prehistoric times, a primitive form of biotechnology was practised by agriculturalists
who established better-quality species of plants and animals by methods of:
 cross-pollination or cross-breeding.
 training and selective breeding of animals,
 the cultivation of crops
 and the utilization of micro-organisms to produce products such as cheese, yogurt,
bread, beer and wine.
 The domestication of animals and plants as a reliable source of food.
 Earliest examples are rice, barley and wheat. Wild animals to produce milk or meat,
production of cheese, yogurt and bread, various alcoholic drinks such as beer and
wine

Definition of Biotechnology

 The simplest way to define biotechnology is to split this word into its two constituent parts
(biotechnology = biology + technology).
 By considering these two key words we can define biotechnology as a set of techniques
that are employed to manipulate living organisms, or utilize biological agents or their
components, to produce useful products/services.
 It is now being used in numerous disciplines including bioremediation, energy production
and food processing agriculture.
 DNA fingerprinting is often practiced in forensics.
 Insulin production and other biotech-based medicines (biopharmaceuticals) are produced
through cloning of vectors with genes of interest (GOIs).
 Immunoassays are frequently utilized in medicine for drug efficiency and pregnancy testing.
In addition, immunoassays are also utilized by farmers to find hazardous levels of
pesticides, herbicides and toxins in crops and animal-based products.
 Biotechnology also has vast scope in agriculture for the production of plants that are
resistant to insects, weeds and plant diseases. This can be achieved by the introduction of
GOIs using genetic engineering.
 ‘Biotechnology means any scientific application that uses biological systems, living
organisms or derivatives thereof, to produce or alter products or processes for particular
use’
 • ‘The utilization of living organisms, systems or processes constitutes biotechnology’
• Based on the Collins English Dictionary definition, biotechnology is the employment of
living organisms, their parts or processes, to develop active and useful products and to
provide services e.g. waste treatment. The term signifies a broad range of processes,
from the use of earthworms as a source of protein to the genetic modification of bacteria
to offer human gene products, e.g. growth hormones.
• According to the Golden Treasury of Science and Technology, biotechnology is a
discipline based on the harnessing of life processes which are controlled for the bulk
production of valuable substances.

• Biotechnology consists of ‘the controlled employment of biological agents,

e.g. micro-organisms or cellular components, for favorable use’
• Biotechnology is the commercial employment of micro-organisms and living plant and
animal cells to create substances or effects beneficial to people. It includes the production
of antibiotics, vitamins, vaccines, plastics, etc
• ‘Bio’ refers to life and ‘technology’ refers to the application of information for
practical use, i.e. the application of living organisms to create or improve a product
• It involves the industrial application of living organisms or their products, which entails the
intentional manipulation of their DNA molecules. It may mean making a living cell execute
a particular task in a predictable and controllable way
• The term biotechnology is occasionally also applied to processes in which micro-
organisms such as yeasts and bacteria are cultured under strictly controlled
environmental conditions. For this reason, fermentation is occasionally called the
oldest form of biotechnology.
• The Universities Press Dictionary of Biology defines biotechnology as ‘the application of
technology to biological processes for industrial, agricultural and medical purposes’
• The Oxford Dictionary of Biology defines biotechnology as ‘the development of
techniques for the application of biological processes to the production of materials of use
in medicine and industry.’
• Hungarian engineer Karl Ereky first coined the term ‘biotechnology’ in 1919, meaning the
production of products from raw materials with the aid of living organisms

In summary, biotechnology is the application of the theory of engineering and biological

science to generate new products from raw materials of biological origin, e.g. vaccines or food’,
or, in other words, it can also be defined as ‘the exploitation of living organism/s or their
product/s to change or improve human health and human surroundings’

B. HISTORY OF BIOTECHNOLOGY

A breakdown of historical events in biotechnology. The development of biotechnology can

be divided into broad stages or categories, including:
1. Ancient biotechnology (8000–4000 BC): Early history as related to food and shelter;
includes domestication of animals.
2. Classical biotechnology (2000 BC; 1800–1900 AD): Built on ancient biotechnology;
fermentation promotes food production and medicine.
 3. Modern biotechnology (1977): Manipulates genetic information in organisms; genetic
engineering; various technologies enable us to improve crop yield and food quality in
agriculture and to produce a broader array of products in industries.
4. Old and new biotechnology: To produce wine, vinegar, curd, leavened bread, etc,
humans began employing microorganisms as early as 5000 BC. Such processes, based on
the natural capabilities of micro-organisms, are commonly considered to be ‘old’
biotechnology.

 During the prehistoric era some civilizations considered fermentation to be a gift from
their gods.
 Scientific evidence for fermentation was first described by Louis Pasteur in the late 1800s.
He demonstrated a theory known as germ theory, presenting the survival of micro-
organisms and their further effects on the process of fermentation.
 Pasteur’s efforts contributed towards several branches of science.
 In earlier times several traditional medicines were used as biotechnology products, such
as;
 honey, which could be used to treat several respiratory ailments and as an ointment for
wounds.
 In China, soybean curds were used to treat boils.
 Ukrainian farmers once used utilized moldy cheese to treat infected wounds.
 In 1928, Alexander Fleming extracted penicillin, the first antibiotic, from mold.
 In the 1920s Chaim Weizmann used Clostridium acetobutylicum for the conversion of
starch into butanol and acetone (the acetone thus produced was used as an essential
component of explosives during World War I)
 The advent of World War II brought the manufacture of penicillin. The production of
antibiotics from micro-organisms became possible when Fleming discovered penicillin,
which was later produced at a large scale from cultures of Penicillium notatum (this
proved useful for the treatment of wounded soldiers during World War II)
 The focus of biotechnology shifted to pharmaceuticals. The Cold War years were ruled by
work on micro-organisms for the preparation for biological products along with antibiotics
and fermentation processes.

 Pre-1800 (Early applications and speculation): Yeast was utilized to prepare beer
(Sumerians and Babylonians).
 4000 BC: In Egypt, a process was discovered to prepare leavened bread by means of yeast.
 420 BC: Greek philosopher Socrates (470–399 BC) hypothesized on the similar
characteristics between parents and their offspring.
 320 BC: Greek philosopher Aristotle (384–322 BC) theorized that all inheritance originates
from the father.
 1630: William Harvey explained that plants and animals are similar in their reproduction, i.e.
they reproduce sexually.
 1660–1675: Marcello Malpighi (1628–1694) investigated blood circulation in capillaries using
a microscope and found that the brain is connected to the spinal cord by bundles of fibers
which form the nervous system
 1673: Antonie van Leeuwenhoek (1632–1723) was the first researcher to explain micro-
organisms such as protozoa and bacteria, and also identify that these micro-organisms play
an active role in fermentation.
 1701: Giacomo Pylarini found that the deliberate administration of smallpox could prevent its
occurrence later in life, especially in children. Later, this procedure was termed ‘vaccination’
and a process that uses cowpox instead of smallpox was established as the most reliable
treatment
 1809: Nicolas Appert invent a technique using heat to can and sterilize food.
 1856: Carl Ludwig discovered a procedure for keeping animal organs alive under in vitro
conditions.
 1859: Charles Darwin (1809–1882) speculated that animal populations adapt their forms to
eventually best utilize the surroundings, a process he described as ‘natural selection’.
 1863: Pasteur discovered the method of pasteurization.
 1865: Mendel (1822–1884) suggested the laws of heredity to the National Scientific Society
(Brunn, Austria) until 1900 when Hugo de Vries, Erich von Tschermak and Carl Correns
supported Mendel’s mechanism of heredity.
 11870: Walther Flemming discovered mitosis.
 1871: During the period 1873–76 interest in DNA research began. DNA was initially derived
from the sperm of trout (found in the river Rhine).
 1881: Koch explained the agar technique for harvesting bacterial colonies on potato slices,
gelatin and agar medium.
 1902: Human genetics is born: Sutton found that chromosomes (paired) contain certain
elements which are transferred from one generation to another.
 1905: X and Y chromosomes related to gender: Edmund Beecher Wilson and Nettie
Stevens shared the same idea of separating X and Y chromosomes for the determination of
sex. They also demonstrated that a single Y chromosome determines maleness, while two
copies of the X chromosome decide femaleness
 1905–1908 William Bateson and R C Punnett, along with other researchers, found that
several genes alter or modify the action of other genes
 1907: Thomas Hunt Morgan started his investigation into utilizing Drosophila fruit flies to
examine heredity, additionally, he discovered mutation theory.
 1909: Wilhelm Johannsen used the word ‘gene’ to mean the carrier/transporter of heredity.
He also coined the terms ‘genotype’ and ‘phenotype’; the genotype is the genetic
composition/establishment of an organism, whereas the phenotype describes the actual
organism or its morphological characteristics, resulting from a blend of the genotype and a
range of external/environmental factors.
 1926: Morgan published The Theory of the Gene.’ This was based on Mendelian genetics
(breeding investigations and optical microscopy).
 1928: Alexander Fleming studied an old culture of bacteria infected with fungal growth and
found that it did not show any bacterial growth in a radius surrounding a piece of mold
(fungi) in a petri dish. This breakthrough gave birth to the antibiotics era or penicillin age.
 1950: Erwin Chargaff discovered that the same levels of adenine and thymine are present in
DNA, as are the same levels of guanine and cytosine. These associations were later
named ‘Chargaff’s rules’.
 1953: The journal Nature published Watson and Crick’s article based on unfolding the
double-helix structure of DNA.
  1957: Central dogma of DNA—how DNA makes a protein. Crick and Gamov studied ‘central
dogma’, demonstrating how DNA functions to construct protein
 1977–present (modern biotechnology) The dawn of biotech: Biotech-based organizations
started focusing more on the applications of genetic engineering.
 1978: Recombinant insulin: Genentech Inc. announced that its laboratory had achieved the
synthesis of human insulin using rDNA technology
 1980: Patents allowed: The US Supreme Court granted that genetically modified living
organisms could be patented.
 1983: Site-directed mutagenesis Eli Lilly obtained a license to make and sell insulin
 1985: During this period genetic fingerprinting stepped into the court room.
 1986: Chiron Corp. obtained FDA approval for the production of the first recombinant vaccine
for hepatitis
 1990: Patents and money: The first gene-based treatment was performed on a four-year-old
girl suffering from an immunological disorder known as adenosine deaminase deficiency
(ADA) deficiency.
 1992: The US Army started taking blood and tissue samples from all new employees as part
of a ‘genetic dog-tag’. This course of action was intended for better identification of soldiers
killed in battle
 1996: The discovery of a gene linked to Parkinson’s disease offered researchers a significant
new chance for the determination of the cause of, and potential treatments for, the
incapacitating neurological disorder.
 1997: Researchers at the Roslin Institute in Scotland announced that they had cloned a
sheep called Dolly from the cell of an adult ewe.
 1998: A group of researchers succeeded in culturing embryonic stem cells. A number of
researchers at Japan’s Kinki University cloned eight identical calves by means of cells
taken from a single adult cow.
 1999: A fatal neurological disease called bovine spongiform encephalopathy (BSE), also
known as mad cow disease, that spread from cattle to humans, was diagnosed by a new
medical diagnostic examination that facilitated the quick detection of BSE/Creutzfeldt–
Jakob disease (CJD).

I. Ancient biotechnology (pre-1800): In the period before the year 1800, some events of
biotechnological developments are as follows:

 Three important basic needs of human civilization are food, clothes and shelter.
 Domestication of food products, formally called ‘agriculture’.
 Humans understood the importance of water, light and other requirements for the optimal
growth of food plants and the domestication of different wild animals
 Domestication of wild animals was the beginning of the applications of animal breeding.
 Evolution of farming led to the development of methods for food preservation and storage.
 Utilization of cold caves or pots (in the form of leather bags and clay jars) to preserve food
for long-term storage began.
 Cheese can be considered to be one of the first direct products (or by-products) of
biotechnology, since it was prepared by adding rennet (an enzyme found in the stomachs of
calves) to sour milk.
 Among the oldest preservatives, vinegar has a significant importance because of its low pH
and potential in preventing the growth of certain microbes, which means it can be used
successfully in food preservation.
 Man started using mules for transportation, carrying loads and farming, before the days of
tractors or trucks.

Some of the processes and products developed in the ancient period are described in table
Domestication • Food supplies were often seasonal. In winter, food supplies could become
quite low.
• People came up with ways of capturing fish and small animals.
• 15 000 years ago, large animals were difficult to catch.
• People may only have had meat when they found a dead animal.
• Domestication most likely began 11 000–12 000 years ago in the Middle
East.
• It involved the adaptation of organisms so they could be cultured.
• Seen by scientists as the beginning of biotechnology.
• Cattle, goats and sheep were the earliest domesticated food animals.
Food • People knew that some foods rotted, while others changed form and
preservation continued to be good to eat.
• Foods stored in a cool cave did not spoil as quickly.
• Foods heated by fire also did not spoil as quickly.
• Immersion in sour liquids prevented food decay.
• Food was stored in bags of leather or jars of clay.
• Fermentation occurs if certain micro-organisms are present, it creates an
acid condition that slows or prevents spoilage.
Cheese • One of the first food products made through biotechnology.
• Strains of bacteria were added to milk, resulting in sour milk.
• An enzyme called ‘rennet’ was added.
• Rennet comes from the lining of the stomachs of calves. It is genetically
engineered today.
• Not all cheese is made from rennet.
• It may have been first developed by nomadic tribes in Asia some 4000 years
ago.
Yeast • Long used in food preparation and preservation. Used in bread baking.
• Produces a gas in the dough causing the dough to rise.
• Used in fermented products such as vinegar.
• Ethanol production require the use of yeast in at least one stage of
production.
Vinegar • Used in pickling.
• Keeps foods from spoiling.
• Juices and extracts from fruits and grains can be fermented.
• Biblical references to wine indicate the use of fermentation some 3000 years
ago.
• Ancient product used to preserve food.
Fermentation • Process in which yeast enzymes chemically change compounds into
alcohol.
• In making vinegar the first product of fermentation is alcohol.
• In ancient times, this likely happened by accident.
• Advancements occurred in the 1800s and early 1900s.
Fermenters • Allowed better control, especially with vinegar.
• New products such as glycerol, acetone and citric acid resulted.

Antibiotics • First drug produced by microbes.

• Used in both human and veterinary medicine.
• Use of fermentation hastened the development of antibiotics.
• Penicillin was developed in the late 1920s.
• Introduced in the 1940s as a drug used to combat bacterial infections.
• Many kinds are available today.
• Limitations in their use keep disease-producing organisms from developing
immunity to antibiotics.
• Some disease organisms are now resistant to certain antibiotics.

II. Classical biotechnology

Classical biotechnology is the second phase of the development of biotechnology. In the
classical era different observations started pouring in, supported by scientific evidence.

These observations made it possible to solve the puzzles of biotechnology. Each and every
observation has made its own contribution in furthering the exploration of new discoveries. The
fundamental idea of the transfer of genetic information from one generation to another forms the
core of biotechnology.

 Information on the transfer of genetic information was first deciphered by Gregor John
Mendel (1822–1884), an Austrian Augustinian monk. Mendel presented his ideas on the
laws of inheritance to the Natural Science Society in Brunn, Austria. He first observed the
transfer of genetic information in a plant, Pisum sativum, commonly known as the pea plant
 Hugo de Vries, Erich von Tschermak and Carl Correns validated Mendel’s discovery work in
1900.
 Fredrich Miescher, a Swiss biologist, reported the existence of nuclein, a compound that
consisted of nucleic acid that he had extracted from pus cells, i.e. white blood cells . During
this time the nucleus in cells was discovered
 Robert Koch (1881), a German physician, first proposed the bacterial propagation method
 Walter Hesse discovered the nutrient agar, the most acceptable and useful medium for
obtaining pure microbial cultures, as well as for their identification
 Heinrich Wilhelm Gottfried von Waldeyer-Hartz, a German scientis, coined the term
‘chromosome’ for an organized structure of DNA and protein present in cells
 Edward Jenner, a British physician, and Louis Pasteur, a French biologist, respectively
developed vaccinations against smallpox and rabies
 The principle of genetics in inheritance was redefined by T H Morgan. He showed
inheritance, and the role of chromosomes in inheritance, using fruit flies (Drosophila
melanogaster).
 Wilhelm Johannsen (1857–1927 coined the term ‘gene. He described the gene as the
carrier of heredity. Afterwards Johannsen coined the terms ‘genotype’ to describe the
genetic constitution of an organism, and ‘phenotype’ to describe the actual organism.\
 Alexander Fleming, a British physician, discovered antibiotics when he observed that one
micro-organism can be used to kill another micro-organism. He discovered penicillin, the
antibacterial toxin from the mold P. notatum, which could be used against many infectious
diseases.
III. Modern biotechnology
A major obstacle to scientific discoveries was the Second World War. After the war, some
essential discoveries were explored. These discoveries form the basis for modern
biotechnology and have brought this field to its current status. Some of the prominent events of
the modern age of biotechnology are highlighted in table

Table 1.3. Historical and current events that form the basis of modern biotechnology.
Year Discoveries
The 1950s
1952 • George Otto Gey created a continuous cell line taken from a human cervical
carcinoma. This cell line, known as HeLa, is still used in therapeutic research.
1953 • Watson and Crick explored DNA as a genetic material, and discovered its structure,
called the double-helix.
1954 • Joseph Murray carried out the first kidney transplant between identical twins.
1960 • A French researcher discovered messenger RNA (mRNA).
The 1970s

1972 • DNA ligases, which join DNA fragments together, were used for the first time.
• The DNA composition of humans was discovered to be 99% similar to that of
chimpanzees and gorillas.
1977 • R Austrian et al at the University of Pennsylvania developed the first vaccine for
pneumonia [65].
1978 • Boyer synthesized the human insulin gene (i.e. a synthetic version of it) and inserted
it into the bacterium E. coli, allowing the bacterium to produce human insulin.
• Louise Brown, the first test-tube baby, was born in the UK. The first vaccine for
meningococcal meningitis was developed.
 The 1980s

1980 • The first automatic gene machine was developed in California.

1981  Baruch Blumberg and Irving Millman developed the first vaccine for hepatitis B.
 Researchers in Switzerland cloned mice.
1982  The first transgenic animals were produced by transforming genes from other
animals into mice.
 The FDA supported the first recombinant protein.
1983
 PCR was identified as the most innovative molecular biology technique.
 The DNA fingerprinting technique was discovered
 The first genetically engineered vaccine was discovered for hepatitis B. The whole
1984
genome of the human immunodeficiency virus (HIV) virus was cloned and
sequenced.
 Genetic fingerprinting stepped into the courtroom.
1985  Genentech became the first biotechnology organization to launch its own
biopharmaceutical product.
 Genetically engineered plants resistant to viruses, insects and bacteria were
fieldtested for the first time.
1981  The Human Genome Project was launched.
 Sir Ian Wilmut cloned an adult sheep and called the cloned sheep ‘Dolly’ [59].
2000  Craig Venter sequenced the human genome; the first publicly accessible genomes
would later be those of James Watson and Ventor.

C. SCOPE AND IMPORTANCE OF BIOTECHNOLOGY

Biotechnology is the science of the controlled application of biological agents for beneficial use.
Since biotechnology is not an independent discipline, its well-known integration with allied fields
such as biochemistry, molecular biology and microbiology facilitates the technological
application of biological agents. Therefore, modern biotechnology has developed as a science
with enormous potential for human welfare in areas ranging from food processing to human
health and environmental protection. The major significance of this field of science in different
fields will be evident from the following examples.
D. BRANCHES OF BIOTECHNOLOGY AND BIOTECH PRODUCTS IN THE
PHILIPPINES, IN ASIA AND WORLDWIDE

I. Biotechnology healthcare and medicine in medicine

The scope of biotechnology in medicine is to utilize techniques in living systems to
produce therapeutic proteins, which are usually called biopharmaceuticals or recombinant
proteins. Products such as monoclonal antibodies, DNA and RNA probes are produced for the
diagnosis of various diseases. Additionally, therapeutic protein-based drugs such as insulin and
interferon have been synthesized with bacteria for the treatment of human diseases.
The use of biotechnology in the field of medicine is also known as ‘red’ biotechnology.
It deals with many major and minor aspects of human life, from making medicines more
effective in terms of cost and efficiency, to tackling one of the most difficult branches of medicine,
curing genetic diseases. Red biotechnology covers various potential medicines for diseases
such as cancer and AIDS. It can be divided into four main areas: biopharmaceuticals, gene
therapy, pharmacogenomics and genetic testing.
The second major field of red biotechnology is gene therapy, which deals with the diagnosis
and treatment of genetic diseases and some other diseases such as cancer.
Pharmacogenomics and genetic testing both use techniques of red biotechnology that are
individual-specific. In pharmacogenomics the genetic information of the individual is derived,
and drugs are developed that can be inserted into that particular individual, whereas in genetic
testing different tests are conducted among family members to determine genetic diseases, sex
and carrier screening. It can also be used in paternity disputes.

II. Biotechnology and the environment

Environmental problems such as pollution control, the depletion of natural resources for non-
renewable energy, conservation of biodiversity, etc, are being dealt with using biotechnology.
For example, bacteria are being utilized for the detoxification of industrial effluents, to combat oil
spills, for treatment of sewage and for biogas production. Biopesticides offer an environmentally
safer alternative to chemical pesticides for control of insect pests and diseases.

III. Industrial biotechnology

Industrial biotechnology was established for the large-scale production of alcohol and antibiotics
by micro-organisms. Currently, various pharmaceutical drugs and chemicals such as lactic acid,
glycerine, etc, are being produced by genetic engineering for better quality and quantity.
Biotechnology has provided us with a very efficient and economical technique for the production
of a variety of biochemicals, e.g. immobilized enzymes. Protein engineering is another important
area where existing proteins and enzymes are remodeled for a specific function or to increase
the efficiency of their function.

IV.Biotechnology in research and agriculture

Currently the potential of plant tissue culture is widely utilized for the rapid and economic
clonal multiplication of fruit and forest trees, for the production of virusfree genetic stock and
planting material, as well as in the creation of novel genetic variations through somaclonal
variation. With the aid of rDNA technology, it has now become possible to produce transgenic
plants with desirable genes such as herbicide resistance, disease resistance, increased shelf
life, etc. Techniques such as molecular breeding have been employed to accelerate the process
of crop improvement. For instance, molecular markers, such as restriction fragment length
polymorphism (RFLP), and simple sequence repeats (SSRs) provide potential tools for the
indirect selection of both qualitative and quantitative traits, and also for studying genotypic
diversity

Biotechnology techniques and other products

1. Bioreactors
In earlier times bioreactors were used for centuries to make wine and beer. The bioreactor is
possibly the most important single piece of equipment used in biotechnology. Bioreactors are
the containers or vessels that allow biological processes to take place under optimum
conditions. These reactors, in controlled environments, will yield a useful substance in large
amounts.

2. Cell fusion
This technique involves the fusion of two cells to make a single cell that contains all the genetic
material of the original cells. So far, this technique has been employed to create new plants by
fusing cells from species that do not naturally hybridize (from a cross-breed) and then
generating whole plants from the fused cells.
3. Liposome-based delivery
Liposomes are microscopic spherical structures that develop when lipids form a suspension in
water. Liposomes have important applications in biotechnology since they may offer novel
means of transporting certain drugs to particular parts of the body across the biological
membranes, e.g. peptides could be encapsulated in liposomes and transported across
biological membranes.

4. Cell or tissue culture

This technique allows the growth and division of individual cells in a bath of sterile, nutritive fluid
which often contains hormones and growth substances. This method is used extensively in
biological laboratories, for example, in cancer research, plant breeding and routine analysis of
chromosome karyotopes.
5. DNA fingerprinting
DNA fingerprinting is a technique that is employed for identifying the components of DNA (the
material of the genes) that are unique to a particular individual. Variations in DNA among
different individuals can be used for identification purposes.

6. Cloning
The method of production of identical animals, plants or micro-organisms from a single
individual is known as cloning. In other words, it is a process by which an organism is derived
from a single parent through non-sexual reproduction. Cloning is gifted in nature to those
organisms that reproduce asexually and produce their own clones, e.g. plants, micro-organisms
and simple animals such as corals.
7. Artificial insemination and ET technology
Artificial insemination allows the artificial introduction of semen into the reproductive tract of a
female animal, and is extensively used in breeding animals, such as sheep and cattle.

8. Stem cell technology

In 1908 Alexander Maksimov coined the term ‘stem cell’. During this period two extensive types
of mammalian stem cells were discovered: embryonic stem cells that are isolated from the inner
cell mass of blastocysts, and adult stem cells that are found in adult tissues.
The classical definition of a stem cell requires that it possesses two properties:
• Self-renewal or the ability to go through numerous cycles of cell division while
maintaining an undifferentiated state.
• Potency or the capacity to differentiate into specialized cell types. Stem cells can be of
the following types:
- Totipotent (in other words, omnipotent) stem cells can differentiate into any kind of
cell type. Such cells can construct a complete, viable, organism. They are produced
from the fusion of an egg and sperm cell.
- Pluripotent stem cells come from totipotent cells and can differentiate into almost all
cells, i.e. cells derived from any of the three germ layers.
Pluripotent adult stem cells are rare and usually small in number but can originate in a
number of tissues including umbilical cord blood. I
- Multipotent stem cells can differentiate into a number of cells, however only into
those of a closely connected family of cells. Multipotent stem cells are also originated
in amniotic fluid.
- Oligopotent stem cells can be distinguished into only a few cells, such as lymphoid
or myeloid stem cells.
- Unipotent cells can offer only one cell type, their own; but they have the property of
self-renewal.

BASIC TECHNIQUES USED IN BIOTECHNOLOGY

Techniques Description

Genetic engineering The use of cellular enzymes to manipulate DNA; transferring

(rDNA) technology DNA between unrelated organisms
Protein engineering Used to improve existing/create novel proteins to make useful products
technology
Anti sense or RNAi Can block or decrease the production of certain proteins
technology
Cell and tissue culture Growing cells/tissues under laboratory conditions to produce an entire
technology organism, or to produce new products
Bio informatics Computational analysis of biological data, e.g. sequence analysis
technology macromolecular structures, high-throughput profiling data analysis

Protein separation and Contour-clamped homogeneous electric field gel electrophoresis

identification techniques Agarose gel electrophoresis
Vertical pulse field gradient electrophoresis
Pulsed field electrophoresis
Polyacrylamide gel electrophoresis
Microarray
Isoelectric focusing
Field inversion gel electrophoresis
Two-dimensional (2D) gel electrophoresis
Blotting techniques Nucleic acid blotting
Southern blot analysis
Protein blotting
Northern blot analysis
Dot blot technique
Autoradiography
Sterilization techniques Steam sterilization
Ultraviolet sterilization
Flame sterilization
Filter sterilization
Dry sterilization
Chemical sterilization Alcohol sterilization
PCR-based techniques Single-nucleotide polymorphism
Targeted PCR and sequencing
SSRs or microsatellites
Sequence-targeted microsatellites
Sequence-related amplification polymorphism
Sequence-characterized amplified regions
Sequence-specified amplified polymorphism
Selective amplification of microsatellite polymorphic loci
Gene transfer Chemical methods
techniques • Calcium phosphate co-precipitation
• Polycation-DMSO technique
• PEG-mediated transformation
• DEAE-dextran procedure

Physical methods
• Ultrasound-mediated gene transformation
• Silicon carbide fiber-mediated transformation
• Microinjection
• Macroinjection
 • Liposome-mediated method
• Electroporation
• Biolistics/particle bombardment/microprojectile
• Virus-mediated gene transfer
• Bacteria-mediated gene transfer
• Agrobacterium-mediated gene transfer
Miscellaneous Protoplast fusion techniques, transposon tagging in heterologous
techniques species, techniques used for single-cell cultures
(Bergmann cell plating technique), immobilization techniques, artificial
seed technology, chromosome elimination techniques, rDNA
technology, spectrophotometry (quantitation, enzyme kinetics), nucleic
acid purification and molecular weight determinations, cell separation
methods, protein separation and quantitation, liquid scintillation (double
label) counting, autoradiography (cellular and gross), restriction
enzyme mapping, gene expression and oligonucleotide synthesis

BIOTECH PRODUCTS IN THE PHILIPPINES,ASIA and WORLDWIDE

Bt Corn: Bacillus thuringiensis (Bt) corn is a genetically modified (GM) crop that is resistant to
pests. In the Philippines, Bt corn has been developed and cultivated to address issues related to
corn crop losses due to pests such as the corn borer. It has been approved for commercial
cultivation by the Philippine government's regulatory bodies. (Reference: Philippine Department
of Agriculture - Bureau of Plant Industry)
Golden Rice: Golden Rice is a genetically modified rice variety enriched with beta-carotene, a
precursor of Vitamin A. It is aimed at addressing Vitamin A deficiency, which is prevalent in
many parts of the world, including the Philippines. Field trials of Golden Rice have been
conducted in the Philippines, and it is awaiting regulatory approval for commercial cultivation.
(Reference: International Rice Research Institute - IRRI)
Pest-Resistant Eggplant (Bt Talong): Bt Talong is a genetically modified eggplant variety
engineered to be resistant to the eggplant fruit and shoot borer (EFSB), a major pest affecting
eggplant production in the Philippines. Like Bt corn, Bt Talong has undergone regulatory
approval and field testing. (Reference: Department of Agriculture - Philippines)
Dengue Vaccine (Dengvaxia): Dengvaxia is a vaccine developed to protect against dengue
fever, a mosquito-borne viral infection that is prevalent in tropical countries like the Philippines.
It is the first dengue vaccine to be licensed and approved for use in several countries, including
the Philippines. However, it's important to note that there have been controversies surrounding
its use, particularly regarding its safety and efficacy. (Reference: Sanofi Pasteur)
Bt Cotton (India): India is one of the largest producers of Bt cotton, a genetically modified
variety of cotton that contains genes from the bacterium Bacillus thuringiensis (Bt), making it
resistant to certain pests. Bt cotton has been commercially cultivated in India since its approval
in 2002. (Reference: Kathage, J., & Qaim, M. (2012). Economic impacts and impact dynamics
of Bt (Bacillus thuringiensis) cotton in India. Proceedings of the National Academy of Sciences,
109(29), 11652-11656.)
CRISPR-Cas9 Gene Editing Technology (Worldwide): CRISPR-Cas9 is a revolutionary gene
editing technology that allows precise modification of DNA sequences in various organisms. It
has numerous applications in biotechnology, including agriculture, medicine, and basic research.
 CRISPR-Cas9 has been used for crop improvement, disease modeling, and potential gene
therapies. (Reference: Doudna, J. A., & Charpentier, E. (2014). The new frontier of genome
engineering with CRISPR-Cas9. Science, 346(6213), 1258096.)
.

BRANCHES OF BIOTECHNOLOGY
The definition of biotechnology can be further divided into different areas of biotechnology by
color, according to their area of research. The different types of biotechnology are grouped into
colors, according to the area of research.

 Red biotechnology: This area includes medical procedures such as utilizing organisms
for the production of novel drugs or employing stem cells to replace/regenerate injured
tissues and possibly regenerate whole organs. It could simply be called medical
biotechnology.
 Green biotechnology: Green biotechnology applies to agriculture and involves such
processes as the development of pest-resistant grains and the accelerated evolution of
disease-resistant animals.
 White biotechnology: White (also called gray) biotechnology involves industrial
processes such as the production of new chemicals or the development of new fuels for
vehicles.
 Yellow biotechnology: researches the production of foods to reduce saturated fat levels,
modify calorie intakes, or supplement vitamins.
 Brown biotechnology: focuses on the treatment of arid and desert soils by studying
species that are highly resistant to saline and dry soils.
 Blue biotechnology: Blue biotechnology, rarely mentioned, encompasses processes in the
marine and aquatic environments, such as controlling the proliferation of noxious water-
borne organisms.
 Gray biotechnology: focuses on the preservation and recovery of natural ecosystems that
have been altered by contaminants.
 Gold biotechnology: related to bioinformatics. It is tasked with the analysis of data in
biological processes (DNA sequences, amino acids, etc.).

 Black biotechnology: This includes all research work on micro-organisms that can be
manipulated to attack human health. Its main activities are related to biological warfare and
bioterrorism.

 Orange biotechnology: disseminates interesting information about biotechnology to attract

future scientists.
 Purple biotechnology: studies all legal aspects related to biotechnology (safety measures,
data protection, patents, etc.).
A distinction is made between ‘non-gene biotechnology’ and ‘gene biotechnology’:
• Non-gene biotechnology: Non-gene biotechnology works with whole cells, tissues or
even individual organisms. Non-gene biotechnology is the more popular practice,
involving plant tissue culture, hybrid seed production, microbial fermentation, production
of hybridoma antibodies and immunochemistry.
• Gene biotechnology: Gene biotechnology deals with genes, the transfer of genes from
one organism to another and genetic engineering.