1. BIOTECHNOLOGY AND ITS
APPLICATIONS
CHAPTER 12

2. WE WILL
LEARN
ABOUT
Biotechnological Applications
in Agriculture
Biotechnological Applications
in Medicine
Transgenic Animals
Ethical Issues

3. BIOTECHNOLOGY PUT TO
WORK!
Biotechnology essentially deals with industrial scale production of
biopharmaceuticals and biologicals. The applications of biotechnology include
therapeutics, diagnostics, genetically modified crops for agriculture, processed
food, bioremediation, waste treatment and energy production.
Biotechnology have the following three critical research areas:
(i) To provide the best catalyst in the form of improved organism, usually a
microbe or pure enzyme.
(ii) To create optimal conditions through engineering for a catalyst to act.
(iii) Downstream processing technologies to purify the protein/organic
compound.

4. BIOTECHNOLOGICAL
APPLICATIONS IN
AGRICULTURE

5. BIOTECH IN
AGRICULTU
RE
(i) Agro-chemical based
agriculture
(ii) Organic agriculture
(iii) Genetically engineered
crop-based agriculture
Agrochemicals (agricultural chemicals, agrichemicals) are the various
chemical products that are used in agriculture. In most cases, the term
agrochemical refers to the broad range of pesticide chemicals,
including insecticide chemicals, herbicide chemicals, fungicide
chemicals, and nematicides chemicals (chemicals used to kill round
worms). The term may also include synthetic fertilizers, hormones, and
other chemical growth agents, as well as concentrated stores of raw
animal manure.
Organic and biotech farming can contribute solutions to various
agricultural problems. “Organic production relies on practices, such as
cultural and biological pest management, that can include IPM and
biological control but excludes the use of synthetic chemicals and GE
organisms. The use of GE organisms can also contribute to sustainable
practices by augmenting and replacing certain conventional practices.
For example, plants can be created that increase water use and
fertilizer efficiencies, that remediate soil contaminants, help reduce
greenhouse gases, and produce higher yields without increasing land
usage, particularly in developing countries.
GM technology enables plant breeders to bring together in one plant
useful genes from a wide range of living sources, not just from within
the crop species or from closely related plants. This powerful tool
allows plant breeders to do faster what they have been doing for years
– generate superior plant varieties.
Although, Global agriculture finds itself engrossed in a heated debate
over genetically modified (GM) crops.

6. GREEN REVOLUTION!!
THERE ARE LIMITATIONS!
Green revolution successfully increased the food production many
folds by usinguse of improved crop varieties as well as better
management practices and use of agrochemicals, fertilizers and
pesticides.
Based on traditional breeding techniques
But-
 For farmers in the developing world, agrochemicals are often too expensive
 Further increase in production is not possible by using these methods.
 We need to minimise the use of fertilisers and chemicals
To overcome these genetically modified crop is used.
We need GM crops!!!

7. GENETICALLY MODIFIED
ORGANISMS

9. PLANTS, BACTERIA, FUNGI AND ANIMALS
WHOSE GENES HAVE BEEN ALTERED BY
MANIPULATION ARE CALLED GENETICALLY
MODIFIED ORGANISMS (GMO).
GM PLANTS HAVE MANY APPLICATIONS-
(i) Made crops more tolerant to abiotic stresses (cold, drought, salt, heat).
(ii) Reduced reliance on chemical pesticides (pest-resistant crops).
(iii) Reduce post harvest losses.
(iv) Increased efficiency of mineral usage by plants
(this prevents early exhaustion of fertility of soil).
(v) Enhanced nutritional value of food, e.g., golden rice, i.e., Vitamin ‘A’
enriched rice.
(vi)GM has been used to create tailor-made plants to supply alternative
resources to industries, in the form of starches, fuels and pharmaceuticals

10. Bt stands for Bacillus thuringiensis which produces Bt toxin.
Bt toxin gene has been cloned from the bacteria and been expressed
in plants to provide resistance to insects without the need for
insecticides; in effect created a bio-pesticide.
Examples are Bt cotton, Bt corn, rice, tomato, potato and soyabean
etc.
Bt, stands out representing approximately 95% of microorganisms
used in biological control of agricultural pests.
Besides the economic aspect and the safety to human health, this
bacterium is the most promising for the production of biopesticides
and plant resistant to insects, associated with environmental
Bacillus thuringiensis and Bt
Crops
Transmission
electron
microscopy
of Bacillus
thuringiensis,
spore (S),
and crystal
(C).

11. (a) Bt cotton is created by using some strains of a
bacterium, Bacillus thuringiensis (Bt is short
form,).
(b) This bacterium produces proteins that kills
certain insects such as lepidopterans (tobacco,
budworm and armyworm), coleopterans (beetles)
and dipterans (flies and mosquitoes).
(c) B. thuringiensis forms protein crystals during a
particular phase of their growth. These crystals
contain a toxic insecticidal protein- Cry toxin
(d) Bt toxin protein exist as inactive protoxins (in
plant), but once an insect ingests the inactive
toxin, it is converted into an active form of toxin
due to the alkaline pH of the gut, which solublises
the crystals.
(e) The activated toxin binds to the surface of
Insect resistant GM Crop- Bt
Cotton
IMPORTAN
T

12. The Bt toxin is coded by cry genes.
The choice of genes depends upon the
crop and the targeted pest, as most Bt
toxins are insect-group specific.
Cry genes ►Cry toxins or Bt
toxins
https://www.ars.usda.gov/ARSUserFiles/oc/np/btcotton/btcott
on.pdf
Gene Controls
cryIAc cotton
bollworms
cryIIAb
cryIAb corn borer

13. https://economictimes.indiatimes.com/

14. BT BRINJAL
FSB-resistant brinjal or Bt brinjal was developed using a transformation process
similar to the one used in the development of Bt cotton, a biotech crop that was
planted on 7.6 million hectares in India in 2008.
Bt brinjal incorporates the cry1Ac gene expressing insecticidal protein to confer
resistance against FSB(fruit shoot borer).
The cry1Ac gene is sourced from the soil bacterium Bacillus thuringiensis (Bt).
When ingested by the FSB larvae, the Bt protein is activated in the insect’s
alkaline gut and binds to the gut wall, which breaks down, allowing the Bt spores
to invade the insect’s body cavity. The FSB larvae die a few days later.
Bt Brinjal was developed by the Maharashtra Hybrid Seeds Company (Mahyco).
The company used a DNA construct containing the cry1Ac gene, a CaMV 35S
promoter and the selectable marker genes nptII and aad, to transform young
cotyledons of brinjal plants.
Not part of syllabus

15. RNA interference (RNAi) is a
biological process in which RNA molecules
inhibit gene expression or translation, by
neutralizing targeted mRNA molecules.
RNAi is present in all eukaryotic
organisms as cellular defence by silencing
of specific mRNA due to complementary
dsRNA molecules that bind to and prevents
translation of the mRNA.
Source of this mRNA ?
-Infection by retroviruses
-Transposons
RNA interference
(RNAi)
No
translation

16. PEST RESISTANT PLANTS
Nematodes like Meloidegyne incognitia infects the roots of tobacco plants and causes
reduction in yield.
STEPS:
The infestation of these nematodes can be prevented by the process of RNA
interference (RNAi).
1. The source of complementary dsRNA may be from (1) an infection by viruses having RNA
genomes or (2) mobile genetic elements (transposons) that replicate through RNA
intermediate.
2. Nematode specific genes were introduced into host plant using Agrobacterium vectors.
3. The introduction of DNA produces both sense and anti-sense RNA in the host cells
4. Two RNAs being complementary to each other formed a double stranded (dsRNA) that
initiated RNAi and thus, silence the specific mRNA of the nematode.
5. The parasite could not survive in a transgenic host expressing specific interfering RNA,
and the plant therefore got itself protected from the parasite

19. OTHER
IMPORTANT
GMOS
GOLDEN RICE : Golden Rice was
genetically modified to provide an
essential nutrient that white rice lacks:
beta-carotene, which is converted into
vitamin A in the body.
FLAVR SAVR TOMATO: This transgenic
tomato was engineered such that it was
no longer able to produce
polygalacturonase (PG), which is an
enzyme involved in fruit softening.
Hence these tomatoes had a long shelf
BETTER NUTRITIVE
QUALITY
BETTER SHELF
LIFE

20. DISADVANTAGES OF GM
CROPS
Environmental Hazards
 Unintended harms to other organisms- eg pollinators, grazers
 Reduced effect of pesticides over time- resistance
Human Health Risk
 Allergies
 Effect on gut microbiota

21. BIOTECHNOLO
GICAL
APPLICATIONS
IN MEDICINE
Genetically Engineered
Insulin
Vaccine Production
Gene Therapy
Molecular Diagnosis

22. Recombinant therapeutics approved for marketing in India.

23. 1. GENETICALLY
ENGINEERED INSULIN
Genetically engineered insulin leads
to sufficient availability of insulin for
the management of adult-onset
diabetes.
(a) Insulin used for diabetes was
earlier extracted from the pancreas
of slaughtered cattle and pigs. This
caused allergy or other reactions in
some patients.
(b) Insulin consists of two short
polypeptide chains, i.e. chain-A and

24. FROM PRO-INSULIN TO
MATURE INSULIN
In mammals, insulin is synthesised as a prohormone
(needs to be processed before it becomes a fully
mature and functional hormone) which contains an
extra stretch called the C-peptide.
C-peptide is not present in the mature insulin and is
removed during maturation into insulin.
Thus, the main challenge for the production of
insulin using rDNA techniques was getting insulin
assembled into a mature form.
Eli Lilly an American company in 1983, prepared two
DNA sequences corresponding to A and B-chains of
human insulin and introduced them in plasmids of
E. coli to produce insulin chains. Chains-A and B
were produced separately, extracted and combined

25. Think about whether insulin
can be orally administered to
diabetic people or not. Why?

26. 2. PRODUCTION OF
VACCINES
Vaccines produced through genetic engineering are called
recombinant vaccines. e.g hepatitis-B.
These are of two types:
 (a) Protein vaccines use of specific protein produced by rDNA in vaccine.
 (b) DNA vaccines use of genetically engineered DNA to be injected as
vaccine to produce an immunological response.
Hepatitis vaccine contains the viral envelope protein, hepatitis-B
surface antigen (HB8 Ag). This gene is isolated from yeast
vectors.
Some protein coding genes isolated from pathogens are also
incorporated and expressed in plants produce antigens and are
also called edible vaccines.

28. VACCINE GENERATIONS
First-Generation Vaccines: Attenuated and inactivated vaccines
Second-Generation Vaccines: The subunit, conjugated, and
recombinant(protein) vaccines are in this generation.
Third generation vaccine: Immunogenic potential administration
of a plasmid containing a gene encoding the antigen, known as
genetic vaccines, is categorized as third generation vaccines

29. 3. GENE
THERAPY
It is a collection of methods that allows
correction of a gene defect that has been
diagnosed in a child or embryo. This
method is applied in a person with a
hereditary disease. In this method, genes
are inserted into a person’s cells and
tissues to treat a disease.
There are two different types of
gene therapy depending on which
types of cells are treated:
Somatic gene therapy: transfer
of a section of DNA to any cell of
the body that doesn’t produce
sperm or eggs. Effects of gene
therapy will not be passed onto
the patient’s children.
Germline gene therapy: transfer
of a section of DNA to cells that
produce eggs or sperm. Effects
of gene therapy will be passed
onto the patient’s children and
subsequent generations.

30. FIRST GENE THERAPY
First gene therapy was given to a
four year old girl with Adenosine
Deaminase (ADA) deficiency by M
Blease and WF Andresco in 1990s.
• ADA is caused due to the deletion
of the gene for adenosine
deaminase.
• In some children, ADA deficiency
can be cured by bone marrow
transplantation and enzyme
replacement therapy, but they are
not completely curable.
Some other diseases that can be
treated by gene therapy are
haemophilia, cystic fibrosis,
In first step of gene therapy, lymphocytes
from the blood of the patient are grown in
a culture outside the body.
• A functional ADA cDNA (using a
retroviral vector) is then introduced into
these lymphocytes, which are subsequently
returned to the patient.
• As these cells are mortal, the patient
requires periodic infusion of such
genetically engineered lymphocytes.
• If the gene isolated from bone marrow
cells producing ADA is introduced into
cells at early embryonic stages, it could be
a permanent cure.

32. 4. MOLECULAR DIAGNOSIS –
“PCR”
Molecular diagnosis helps to solve the problem of early diagnosis and treatment of diseases.
(a) Symptoms of disease appear only when the concentration of pathogen get increased
significantly. Using conventional methods of diagnosis (serum and urine analysis), early
detection of diseases is not possible.
(b) To overcome this problem, some molecular diagnosis techniques were developed that
provide early detection of diseases.
►Polymerase Chain Reaction (PCR) helps in early detection of diseases or pathogens
by the amplification of their nucleic acid.
Low concentration of pathogens (bacteria, viruses, etc) in the blood does not allow
its detection. PCR can amplify nucleic acids of such pathogens even when their
concentration is very low.
PCR technique can be used for detecting HIV in suspected AIDS patients, genetic
mutation in suspected cancer patients and in identifying genetic disorders.
► Enzyme Linked Immuno-sorbent Assay (ELISA)
ELISA is based on the principle of antigen-antibody interaction. Infection by
pathogen can be detected by the presence of antigens like proteins, glycoproteins
etc. or by detecting the antibodies synthesised against the pathogen.

33. AFTER PCR: HYBRIDIZATION
WITH LABELED PROBE AND
AUTORADIOGRAPHY
A single stranded DNA or RNA, tagged with a
radioactive molecule is called a probe
Probe is allowed to hybridize to its complementary
DNA in a clone of cells.
This is followed by detection using autoradiography.
The clone having the mutated gene will hence not
appear on the photographic film, because the probe
will not have complementarity with the mutated gene.
MOLECULAR DIAGNOSIS = PCR > HYBRIDIZATION >
AUTORADIOGRAPHY

34. Gene
1
Gene
2
Gene
3
Gene
4
Gene
5
Person
A
Person
B
Person
C
Control
(All normal)
Gene
6
Person A had Gene 2 and Gene 6 mutated

35. PCR
Hybridization
with labelled
probe
http://bio-
Or cancerous
cell

37. ELISA
PRINCIPLE: antigen-antibody interaction.
Infection by pathogen can be detected
(1) by the presence of antigens (proteins, glycoproteins, etc.)
OR
(2) by detecting the antibodies synthesised against the pathogen.
ENZYME
LINKED
IMMUNO
SORBENT
ASSAY

38. TRANSGENIC
ANIMALS
Animals that have had
their DNA manipulated
to possess and express
a foreign gene are
known as transgenic
animals. Transgenic
mice, rats, rabbits,
pigs, sheep, cows and
fish have been
produced.

39. 1. NORMAL PHYSIOLOGY AND
DEVELOPMENT
Transgenic animals can be specifically designed to allow the
study of
How genes are regulated
How they affect the normal functions of the body and its development
e.g., study of complex factors involved in growth such as insulin-like
growth factor.
HOW?
This can be done by introducing genes from other species that alter the
formation of this factor and studying the biological effects that result.
Information is obtained about the biological role of the factor in the
body.

40. 2. STUDY OF DISEASE
Studies on transgenic models have provided new insights into
the genetic origin of certain diseases and have improved our
understanding of pathological processes on the cellular level.
These are specially made to serve as models for human diseases
so that investigation of new treatments for diseases is made
possible.
Today transgenic models exist for many human diseases such
as cancer, cystic fibrosis, rheumatoid arthritis and Alzheimer’s.
Transgenic animals have also been used to study aspects of
tumor development, immune regulation etc

41. 3. BIOLOGICAL PRODUCTS
Medicines required to treat certain human
diseases can contain biological products, but
such products are often expensive to make.
Transgenic animals that produce useful
biological products can be created by the
introduction of the portion of DNA (or genes)
which codes for the desired product
Eg- Human protein (α-1-antitrypsin) used to
treat emphysema. Similar attempts are being
made for treatment of phenylketonuria (PKU) and
cystic fibrosis.
►In 1997, first transgenic cow, Rosie, produced
human protein-enriched milk (2.4 grams per
litre). The milk contained the human alpha-

42. 4. VACCINE SAFETY
Transgenic mice are being developed
for use in testing the safety of
vaccines before they are used on
humans.
Transgenic mice are used to test the
safety of the polio vaccine.
If successful and found to be reliable,
they could replace the use of monkeys
to test the safety of batches of the
vaccine.
Monkey is used because only primates
are naturally susceptibleto poliovirus

43. 5. CHEMICAL SAFETY
TESTING
This is known as toxicity/safety testing. The procedure is
the same as that used for testing toxicity of drugs.
Transgenic animals are made that carry genes which
make them more sensitive to toxic substances than non-
transgenic animals.
They are then exposed to the toxic substances and the
effects studied. Toxicity testing in such animals will allow
us to obtain results in less time.

44. STEM CELL
TECHNOLOGY

45. STEM CELLS
“Stem cells are special undifferentiated human cells that can develop into many
different types of cells, from muscle cells to brain cells.”
Stem cells also have the ability to repair the damaged cells. These cells have strong
healing power. They can evolve into any types of cell.
Researches are going on and it is believed that stem cell therapies can cure ailments
like paralysis and Alzheimer’s as well.
Stem cells are of the following different types:
Embryonic
Stem Cells
Adult Stem
Cells
Induced
Pluripotent
Stem Cells
Mesenchymal
stem cells

46. SOURCE OF STEM CELL
Stem Cells originate from different parts of the body.
Adult stem cells can be found in specific tissues in the human
body (eg bone marrow) . Matured cells are specialized to conduct
various functions. Generally, these cells can develop the kind of
cells found in tissues where they reside.
Embryonic Stem Cells are derived from 5-day old blastocysts that
develop into embryos and are pluripotent in nature. These cells
can develop any type of cell and tissue in the body. These cells
have the potential to regenerate all the cells and tissues that have
been lost because of any kind of injury or disease.

48. APPLICATIONS OF STEM
CELLS
Tissue Regeneration: This is the most important application of stem cells. The stem cells can be used to
grow a specific type of tissue or organ.
Treatment of Cardiovascular Disease: A team of researchers have developed blood vessels in mice using
human stem cells.
Treatment of Brain Diseases: Stem cells can also treat diseases such as Parkinson’s disease and Alzheimer’s.
These can help to replenish the damaged brain cells.
Blood Disease Treatment: The adult hematopoietic stem cells are used to treat cancers, sickle cell anaemia,
and other immunodeficiency diseases. These stem cells can be used to produce red blood cells and white
blood cells in the body.

49. FEW CONTRIBUTIONS OF
BIOTECHNOLOGY TO THE INDUSTRY
Single cell proteins (from bacteria, yeast, algae) food supplements
Fuel (ethanol, biogas)
Mineral extraction
Immobilization of enzymes
Engineered enzymes
Production of immunotxoins
Production of antibiotics

50. ETHICAL
ISSUES

51. GENETIC
ENGINEERING
APPRAISAL
COMMITTEE
(GEAC)
The GEAC is India’s apex biotechnology
regulatory body which makes decisions
regarding the validity of GM research and the
safety of introducing GM-organisms for public
services.
ROLE OF GEAC: It regulates the use,
manufacture, storage, export and import of
hazardous microorganisms or genetically-
engineered organisms and cells in India.
Biopatent: A patent is the right granted by a
government to an inventor to prevent others
from making commercial use of his invention.
Now, patents are granted for biological entities
and for products derived from biological
resources.
The Indian Parliament has recently cleared the
second amendment of the Indian Patents Bill,
that takes such issues, including patent terms.
emergency provisions into consideration and
research and development initiative.

52. BIOPIRACY IS THE TERM
USED TO REFER TO THE
USE OF BIO-RESOURCES
BY
MULTINATIONAL
COMPANIES AND OTHER
ORGANISATIONS WITHOUT
PROPER
AUTHORISATION FROM THE
COUNTRIES AND PEOPLE
CONCERNED WITHOUT
COMPENSATORY PAYMENT
BIOPIRAC
Y

53. INDIAN PATENT
BATTLES
In 1997, an American company RiceTec got patent rights on Basmati rice
through the US Patent and Trademark Office.
This allowed the company to sell a ‘new variety of Basmati, in the US and
abroad.
This ‘new’ variety of Basmati had actually been derived from Indian farmer’s
varieties. Indian Basmati was crossed with semi-dwarf varieties and claimed
as an invention or a novelty.
Several attempts have also been made to patent uses, products and
processes based on Indian traditional herbal medicines, e.g., turmeric and
neem.
READ: https://www.mondaq.com/india/patent/586384/traditional-knowledge-and-patent-issues-an-overview-of-turmeric-
basmati-neem-cases
Rice is an important food grain, the presence of which goes
back thousands of years in Asia’s agricultural history. There
are an estimated 200,000 varieties of rice in India alone. The
diversity of rice in India is one of the richest in the world.
Basmati rice is distinct for its unique aroma and flavour and
27 documented varieties of Basmati are grown in India.

54. https://www.nature.com/articles/35698

56. TRADITIONAL KNOWLEDGE
Most of the industrialized nations are rich
financially but poor in biodiversity and
traditional knowledge.
In contrast the developing and the
underdeveloped world is rich in biodiversity
and traditional knowledge related to bio-
resources.
Traditional knowledge related to bio-
resources can be exploited to develop modern
applications and can also be used to save
time, effort and expenditure during their
Traditional
Knowledge
(TK) is a living
body of
knowledge
that is
developed,
sustained and
passed on
from
generation to
generation
within a

57. INDIAN PATENTS BILL
Source: https://ipindia.gov.in/history-of-indian-patent-
system.htm

58. THANKYOU
© BHOOMIKA

59. WORKSHEET
1. Gene expression can be controlled with the help of RNA. Explain the method with an
example.
2. Highlight any four areas where genetic modification of plants has been useful.
3. Discuss briefly how a probe is used in molecular diagnostics.
4. How is a mature, functional insulin hormone different from its prohormone form? List
the disadvantages of insulin obtained from the pancreas of slaughtered cows and pigs.
List the advantages of recombinant insulin.
OR
(How was Insulin obtained before the advent of rDNA technology? What were the
problems encountered?)
5. You have identified a useful gene in bacteria. Make a flow chart of the steps that you
would follow to transfer this gene to a plant.
6. Diagrammatically represent the experimental steps in cloning and expressing an human
gene (say the gene for growth hormone) into a bacterium like E. coli ?
7. What are transgenic bacteria? Illustrate using any one example.
8. Find out from internet what is golden rice.
9. Consult internet and find out how to make orally active protein pharmaceutical. What is