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amrit_rj
BIOTECHNOLOGY
● Biotechnology, deals with industrial scale
production of biopharmaceuticals and
biologicals using genetically modified
microbes, fungi, plants and animals.
● The applications of biotechnology include
therapeutics, diagnostics, genetically modified
crops for agriculture processed food,
bioremediation, waste treatment, and energy
production.
amrit_rj
Three critical research areas of
biotechnology are:
Providing the best catalyst in the form of
improved organism usually a microbe or pure
enzyme.
Creating optimal conditions through
engineering for a catalyst to act.
Downstream processing technologies to
purify the protein/organic compound.
1.
2.
3.
amrit_rj
BIOTECHNOLOGICAL APPLICATIONS IN
AGRICULTURE
amrit_rj
Organic agriculture
2.
Agro-chemical based
agriculture
1.
Genetically engineered
crop-based agriculture
3. Options for
increasing
food
production
amrit_rj
GREEN REVOLUTION
● The Green Revolution succeeded in
tripling the food supply but yet it was not
enough to feed the growing human
population.
● Increased yields have partly been due to
the use of improved crop varieties, but
mainly due to the use of better
management practices and use of
agrochemicals (fertilisers and pesticides).
amrit_rj
GENETICALLY MODIFIED FOOD
● Plants, bacteria, fungi and animals whose genes have been altered by manipulation
are called Genetically Modified Organisms (GMO).
● GM plants have been useful in many ways. Genetic modification has:
made crops more tolerant to abiotic stresses (cold, drought, salt,
heat).
reduced reliance on chemical pesticides (pest-resistant crops).
helped to reduce post harvest losses.
1.
2.
3.
increased efficiency of mineral usage by plants (this prevents early
exhaustion of fertility of soil).
enhanced nutritional value of food, e.g., golden rice, i.e., Vitamin ‘A’
enriched rice.
5.
4.
amrit_rj
List any four ways by which GMO’s have been useful
for enhanced crop output. (3 marks, 2019)
amrit_rj
List any four ways by which GMO’s have been useful for enhanced
crop output. (3 marks, 2019)
(i) genetically modified crops are more tolerant to abiotic stresses (cold, drought,
salt, heat
(ii) These genetic modifications have helped to reduce post harvest losses implying
less damage to crop yield.
(iii) Genetic modification has reduced reliance on chemical pesticides by creating
pest resistant crops. eg. Tobacco, leading to greater crop yield.
(iv) Creation of insect resistant crops such as Bt cotton and Bt corn has enhanced
crop yield.
(v) Increased efficiency of mineral usage by GM plants prevents early exhaustion of
fertility of soil resulting in increased yield
amrit_rj
Bt COTTON
● Bt toxin gene (cry gene) has been cloned
from the bacteria and been expressed in
plants to provide resistance to insects.
● Bt toxin kills lepidopterans (tobacco
budworm, armyworm), coleopterans
(beetles) and dipterans (flies, mosquitoes).
● B. thuringiensis forms protein crystals
(insecticidal protein) during a particular
phase of their growth.
amrit_rj
Mechanisms of Action
Bt toxin exists in inactive form (Protoxin)
Ingested by the bite of insect
Alkaline conditions of insect gut activate toxin.
Active toxin binds to midgut epithelial cells
Pores are created by swelling & lysis - Insect Death
amrit_rj
Bt COTTON
● The choice of genes depends upon the crop and the targeted pest, as
most Bt toxins are insect-group specific.
● The toxin is coded by a gene cryIAc named cry.
● There are a number of them, for example, the proteins encoded by the
genes:
(i) cryIAc and cryIIAb - control the cotton bollworms
(ii) cryIAb - controls corn borer.
amrit_rj
What are Cry genes? In which organism are they present ?
( 1 mark, 2017)
amrit_rj
What are Cry genes? In which organism are they present ? ( 1 mark, 2017)
Cry genes codes for a toxin which is poisonous to some insects thus
giving resistant to the plants. They are present in bacterium Bacillus
thuriengiensis.
amrit_rj
When Bacillus thuringiensis enters a certain insect’s body, the
insect gets killed, but itself remains unaffected. Explain how it is
possible. (3 marks, 2020)
amrit_rj
When Bacillus thuringiensis enters a certain insect’s body, the
insect gets killed, but itself remains unaffected. Explain how it is
possible. (3 marks, 2020)
Bt is a microbe that lives in the soil. It produces proteins that are poisonous to young
insects (larvae). Bt comes in a variety of forms. Each one is aimed at a different type
of insect. Beetles, mosquitoes, black flies, caterpillars, and moths are among the
insects targeted.
1) Routine testing is required with Bt pesticides to ensure that unwanted toxins and
microbes are not present. Since 1961, the US Environmental Protection Agency
(EPA) has approved Bt for use in pesticides.
2) When Bacillus thuringiensis enters a certain insect’s body, the insect gets killed,
but itself remains unaffected because it contains some acids that produce protein
from substances that are toxic to insects.
amrit_rj
PEST RESISTANT PLANTS
● Several nematodes parasitise a wide variety of
plants and animals including human beings.
● A nematode Meloidogyne incognita infects the
roots of tobacco plants and causes a great
reduction in yield.
● A novel strategy was adopted to prevent this
infestation which was based on the process of
RNA interference (RNAi).
● RNAi takes place in all eukaryotic organisms as
a method of cellular defense.
amrit_rj
STEPS IN RNAi
(i) dsRNA is used to silence target RNA.
(ii) Source of dsRNA can be retroviruses or transposons.
(iii) Using Agrobacterium vectors, nematode specific genes were introduced into the
host plant.
(iv) The introduction of DNA was such that it produced both sense and antisense
RNA in the host cells.
(v) These two RNA’s being complementary to each other formed a double stranded
(dsRNA) that initiated RNAi and thus, silenced the specific mRNA of the
nematode.
(vi) The consequence was that the parasite could not survive in a transgenic host
expressing specific interfering RNA.
(vii) The transgenic plant therefore got itself protected from the parasite.
amrit_rj
How has the use of Agrobacterium as vectors helped in
controlling Meloidegyne incognitia infestation in tobacco
plants ? Explain in correct sequence. ( 3 marks, 2018)
amrit_rj
How has the use of Agrobacterium as vectors helped in controlling Meloidegyne
incognitia infestation in tobacco plants ? Explain in correct sequence. ( 3 marks, 2018)
Using Agrobacterium vector nematode specific genes introduced into host
plant
- Sense and antisense strands of mRNA are produced
- ds RNA is formed
- ds RNA initiates RNAi
- Prevents translation of mRNA / silencing of mRNA of parasite / nematode
- Parasite will not survive
amrit_rj
BIOTECHNOLOGICAL APPLICATIONS IN
MEDICINE
amrit_rj
● Insulin used for diabetes was earlier extracted from
pancreas of slaughtered cattle and pigs.
● Insulin from an animal source, though caused some
patients to develop allergy or other types of reactions
to the foreign protein.
● Insulin consists of two short polypeptide chains:
chain A and chain B, that are linked together by
disulphide bridges .
● In mammals, including insulin is synthesised as a
prohormone which contains an extra stretch called
the C peptide.
● This C peptide is not present in the mature insulin.
GENETICALLY ENGINEERED INSULIN
amrit_rj
● In 1983, Eli Lilly an American
company 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.
GENETICALLY ENGINEERED INSULIN
amrit_rj
Mention the chemical change that pro-insulin undergoes, to
be able to act as mature insulin. ( 1 mark, 2018)
amrit_rj
Mention the chemical change that pro-insulin undergoes, to be
able to act as mature insulin. ( 1 mark, 2018)
Proinsulin is a polypetide that consists of the 'A' subunit and 'B'
subunit of the insulin joined together by a 'C' chain. The proinsulin
undergoes a chemical change where in the 'C' peptide is cleaved
and removed. The removal is essential for proinsulin to be able to
function as mature insulin
amrit_rj
Explain the various steps involved in the production of
artificial insulin? ( 3 marks , 2017)
amrit_rj
Explain the various steps involved in the production of artificial insulin? ( 3
marks , 2017)
Genetically Engineered Insulin: -
* Insulin contains two short polypeptide chains: chain A and chain B linked together by
disulphide bridges.
* In mammals insulin is synthesised as a pro-hormone. It contains an extra stretch called
C-peptide.
* C-peptide is absent in the mature insulin and is removed during maturation into insulin.
* Production of insulin by rDNA techniques was achieved by an American company, Eli
Lilly in 1983.
● It prepared two DNA sequences corresponding to A and B, chains of human insulin
and introduced them in plasmids of E. coli for production. The A and B chains
produced were separated, extracted and combined, by creating disulfide bonds to
form human insulin.
amrit_rj
● Gene therapy is a collection of methods that allows correction of a gene defect
that has been diagnosed in a child/embryo.
● Here genes are inserted into a person’s cells and tissues to treat a disease.
● The first clinical gene therapy was given in 1990 to a 4-year old girl with
adenosine deaminase (ADA) deficiency.
● The disorder is caused due to the deletion of the gene for adenosine
deaminase.
● In some children ADA deficiency can be cured by bone marrow
transplantation; in others it can be treated by enzyme replacement therapy.
● In others it can be treated by enzyme replacement therapy, in which functional
ADA is given to the patient by injection.
● But the problem with both of these approaches that they are not completely
curative.
GENE THERAPY
amrit_rj
● As a first step towards 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.
● However, as these cells are not immortal, the patient requires
periodic infusion of such genetically engineered lymphocytes.
● However, if the gene isolate from marrow cells producing
ADA is introduced into cells at early embryonic stages, it
could be a permanent cure.
GENE THERAPY
amrit_rj
BIOTECHNOLOGICAL APPLICATIONS IN
MEDICINE
Polymerase Chain
Reaction
ELISA
amrit_rj
● PCR is now routinely used to detect HIV in suspected AIDS patients.
● Used to detect mutations in genes in suspected cancer patients too.
● It is a powerful technique to identify many other genetic disorders
● A single stranded DNA or RNA, tagged with a radioactive molecule (probe) is
allowed to hybridise to its complementary DNA in a clone of cells
● 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.
Polymerase Chain Reaction
amrit_rj
● Principle: Antigen-antibody
interaction.
● Infection by pathogen can be detected
by the presence of antigens (proteins,
glycoproteins, etc.)
● By detecting the antibodies
synthesised against the pathogen.
ELISA
amrit_rj
Normal physiology and development
Study of disease
Biological products
Vaccine safety
TRANSGENIC ANIMALS
Chemical safety testing
amrit_rj
Bioethical issues
● Some ethical standards are required to
evaluate the morality of all human activities
that might help or harm living organisms.
● Genetic modification of organisms can have
unpredictable results when such organisms
are introduced into the ecosystem.
● Therefore, the Indian Government has set up
organisations such as GEAC (Genetic
Engineering Approval Committee), which will
make decisions regarding the validity of GM
research and the safety of introducing
GM-organisms for public services.
amrit_rj
BIOPIRACY
Use of bio-resources by multinational companies
and other organisations without proper authorisation
from the countries and people concerned without
compensatory payment
Indian Parliament has recently cleared the second
amendment of the Indian Patents Bill, that takes
such issues into consideration, including
● Patent terms emergency provisions
● Research and development initiative.
amrit_rj
What is biopiracy ? ( 1 mark, 2016)
amrit_rj
What is biopiracy ? ( 1 mark, 2016)
Biopiracy is the use of bioresources by multinational companies and
other organization without proper authorization/compensation
payment to the concern country /organisation.
Please find the DPP V-Quiz
link in the description box
Course sale price Rs.
2700/-(after discount)
NEET Crash Course-Lite
Recordings will be
available till your NEET
examination
9th May,2022
ARAPRO
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Biotechnology&+its+applications.pdf

  • 2. amrit_rj BIOTECHNOLOGY ● Biotechnology, deals with industrial scale production of biopharmaceuticals and biologicals using genetically modified microbes, fungi, plants and animals. ● The applications of biotechnology include therapeutics, diagnostics, genetically modified crops for agriculture processed food, bioremediation, waste treatment, and energy production.
  • 3. amrit_rj Three critical research areas of biotechnology are: Providing the best catalyst in the form of improved organism usually a microbe or pure enzyme. Creating optimal conditions through engineering for a catalyst to act. Downstream processing technologies to purify the protein/organic compound. 1. 2. 3.
  • 5. amrit_rj Organic agriculture 2. Agro-chemical based agriculture 1. Genetically engineered crop-based agriculture 3. Options for increasing food production
  • 6. amrit_rj GREEN REVOLUTION ● The Green Revolution succeeded in tripling the food supply but yet it was not enough to feed the growing human population. ● Increased yields have partly been due to the use of improved crop varieties, but mainly due to the use of better management practices and use of agrochemicals (fertilisers and pesticides).
  • 7. amrit_rj GENETICALLY MODIFIED FOOD ● Plants, bacteria, fungi and animals whose genes have been altered by manipulation are called Genetically Modified Organisms (GMO). ● GM plants have been useful in many ways. Genetic modification has: made crops more tolerant to abiotic stresses (cold, drought, salt, heat). reduced reliance on chemical pesticides (pest-resistant crops). helped to reduce post harvest losses. 1. 2. 3. increased efficiency of mineral usage by plants (this prevents early exhaustion of fertility of soil). enhanced nutritional value of food, e.g., golden rice, i.e., Vitamin ‘A’ enriched rice. 5. 4.
  • 8. amrit_rj List any four ways by which GMO’s have been useful for enhanced crop output. (3 marks, 2019)
  • 9. amrit_rj List any four ways by which GMO’s have been useful for enhanced crop output. (3 marks, 2019) (i) genetically modified crops are more tolerant to abiotic stresses (cold, drought, salt, heat (ii) These genetic modifications have helped to reduce post harvest losses implying less damage to crop yield. (iii) Genetic modification has reduced reliance on chemical pesticides by creating pest resistant crops. eg. Tobacco, leading to greater crop yield. (iv) Creation of insect resistant crops such as Bt cotton and Bt corn has enhanced crop yield. (v) Increased efficiency of mineral usage by GM plants prevents early exhaustion of fertility of soil resulting in increased yield
  • 10. amrit_rj Bt COTTON ● Bt toxin gene (cry gene) has been cloned from the bacteria and been expressed in plants to provide resistance to insects. ● Bt toxin kills lepidopterans (tobacco budworm, armyworm), coleopterans (beetles) and dipterans (flies, mosquitoes). ● B. thuringiensis forms protein crystals (insecticidal protein) during a particular phase of their growth.
  • 11. amrit_rj Mechanisms of Action Bt toxin exists in inactive form (Protoxin) Ingested by the bite of insect Alkaline conditions of insect gut activate toxin. Active toxin binds to midgut epithelial cells Pores are created by swelling & lysis - Insect Death
  • 12. amrit_rj Bt COTTON ● The choice of genes depends upon the crop and the targeted pest, as most Bt toxins are insect-group specific. ● The toxin is coded by a gene cryIAc named cry. ● There are a number of them, for example, the proteins encoded by the genes: (i) cryIAc and cryIIAb - control the cotton bollworms (ii) cryIAb - controls corn borer.
  • 13. amrit_rj What are Cry genes? In which organism are they present ? ( 1 mark, 2017)
  • 14. amrit_rj What are Cry genes? In which organism are they present ? ( 1 mark, 2017) Cry genes codes for a toxin which is poisonous to some insects thus giving resistant to the plants. They are present in bacterium Bacillus thuriengiensis.
  • 15. amrit_rj When Bacillus thuringiensis enters a certain insect’s body, the insect gets killed, but itself remains unaffected. Explain how it is possible. (3 marks, 2020)
  • 16. amrit_rj When Bacillus thuringiensis enters a certain insect’s body, the insect gets killed, but itself remains unaffected. Explain how it is possible. (3 marks, 2020) Bt is a microbe that lives in the soil. It produces proteins that are poisonous to young insects (larvae). Bt comes in a variety of forms. Each one is aimed at a different type of insect. Beetles, mosquitoes, black flies, caterpillars, and moths are among the insects targeted. 1) Routine testing is required with Bt pesticides to ensure that unwanted toxins and microbes are not present. Since 1961, the US Environmental Protection Agency (EPA) has approved Bt for use in pesticides. 2) When Bacillus thuringiensis enters a certain insect’s body, the insect gets killed, but itself remains unaffected because it contains some acids that produce protein from substances that are toxic to insects.
  • 17. amrit_rj PEST RESISTANT PLANTS ● Several nematodes parasitise a wide variety of plants and animals including human beings. ● A nematode Meloidogyne incognita infects the roots of tobacco plants and causes a great reduction in yield. ● A novel strategy was adopted to prevent this infestation which was based on the process of RNA interference (RNAi). ● RNAi takes place in all eukaryotic organisms as a method of cellular defense.
  • 18. amrit_rj STEPS IN RNAi (i) dsRNA is used to silence target RNA. (ii) Source of dsRNA can be retroviruses or transposons. (iii) Using Agrobacterium vectors, nematode specific genes were introduced into the host plant. (iv) The introduction of DNA was such that it produced both sense and antisense RNA in the host cells. (v) These two RNA’s being complementary to each other formed a double stranded (dsRNA) that initiated RNAi and thus, silenced the specific mRNA of the nematode. (vi) The consequence was that the parasite could not survive in a transgenic host expressing specific interfering RNA. (vii) The transgenic plant therefore got itself protected from the parasite.
  • 19. amrit_rj How has the use of Agrobacterium as vectors helped in controlling Meloidegyne incognitia infestation in tobacco plants ? Explain in correct sequence. ( 3 marks, 2018)
  • 20. amrit_rj How has the use of Agrobacterium as vectors helped in controlling Meloidegyne incognitia infestation in tobacco plants ? Explain in correct sequence. ( 3 marks, 2018) Using Agrobacterium vector nematode specific genes introduced into host plant - Sense and antisense strands of mRNA are produced - ds RNA is formed - ds RNA initiates RNAi - Prevents translation of mRNA / silencing of mRNA of parasite / nematode - Parasite will not survive
  • 22. amrit_rj ● Insulin used for diabetes was earlier extracted from pancreas of slaughtered cattle and pigs. ● Insulin from an animal source, though caused some patients to develop allergy or other types of reactions to the foreign protein. ● Insulin consists of two short polypeptide chains: chain A and chain B, that are linked together by disulphide bridges . ● In mammals, including insulin is synthesised as a prohormone which contains an extra stretch called the C peptide. ● This C peptide is not present in the mature insulin. GENETICALLY ENGINEERED INSULIN
  • 23. amrit_rj ● In 1983, Eli Lilly an American company 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. GENETICALLY ENGINEERED INSULIN
  • 24. amrit_rj Mention the chemical change that pro-insulin undergoes, to be able to act as mature insulin. ( 1 mark, 2018)
  • 25. amrit_rj Mention the chemical change that pro-insulin undergoes, to be able to act as mature insulin. ( 1 mark, 2018) Proinsulin is a polypetide that consists of the 'A' subunit and 'B' subunit of the insulin joined together by a 'C' chain. The proinsulin undergoes a chemical change where in the 'C' peptide is cleaved and removed. The removal is essential for proinsulin to be able to function as mature insulin
  • 26. amrit_rj Explain the various steps involved in the production of artificial insulin? ( 3 marks , 2017)
  • 27. amrit_rj Explain the various steps involved in the production of artificial insulin? ( 3 marks , 2017) Genetically Engineered Insulin: - * Insulin contains two short polypeptide chains: chain A and chain B linked together by disulphide bridges. * In mammals insulin is synthesised as a pro-hormone. It contains an extra stretch called C-peptide. * C-peptide is absent in the mature insulin and is removed during maturation into insulin. * Production of insulin by rDNA techniques was achieved by an American company, Eli Lilly in 1983. ● It prepared two DNA sequences corresponding to A and B, chains of human insulin and introduced them in plasmids of E. coli for production. The A and B chains produced were separated, extracted and combined, by creating disulfide bonds to form human insulin.
  • 28. amrit_rj ● Gene therapy is a collection of methods that allows correction of a gene defect that has been diagnosed in a child/embryo. ● Here genes are inserted into a person’s cells and tissues to treat a disease. ● The first clinical gene therapy was given in 1990 to a 4-year old girl with adenosine deaminase (ADA) deficiency. ● The disorder is caused due to the deletion of the gene for adenosine deaminase. ● In some children ADA deficiency can be cured by bone marrow transplantation; in others it can be treated by enzyme replacement therapy. ● In others it can be treated by enzyme replacement therapy, in which functional ADA is given to the patient by injection. ● But the problem with both of these approaches that they are not completely curative. GENE THERAPY
  • 29. amrit_rj ● As a first step towards 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. ● However, as these cells are not immortal, the patient requires periodic infusion of such genetically engineered lymphocytes. ● However, if the gene isolate from marrow cells producing ADA is introduced into cells at early embryonic stages, it could be a permanent cure. GENE THERAPY
  • 31. amrit_rj ● PCR is now routinely used to detect HIV in suspected AIDS patients. ● Used to detect mutations in genes in suspected cancer patients too. ● It is a powerful technique to identify many other genetic disorders ● A single stranded DNA or RNA, tagged with a radioactive molecule (probe) is allowed to hybridise to its complementary DNA in a clone of cells ● 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. Polymerase Chain Reaction
  • 32. amrit_rj ● Principle: Antigen-antibody interaction. ● Infection by pathogen can be detected by the presence of antigens (proteins, glycoproteins, etc.) ● By detecting the antibodies synthesised against the pathogen. ELISA
  • 33. amrit_rj Normal physiology and development Study of disease Biological products Vaccine safety TRANSGENIC ANIMALS Chemical safety testing
  • 34. amrit_rj Bioethical issues ● Some ethical standards are required to evaluate the morality of all human activities that might help or harm living organisms. ● Genetic modification of organisms can have unpredictable results when such organisms are introduced into the ecosystem. ● Therefore, the Indian Government has set up organisations such as GEAC (Genetic Engineering Approval Committee), which will make decisions regarding the validity of GM research and the safety of introducing GM-organisms for public services.
  • 35. amrit_rj BIOPIRACY Use of bio-resources by multinational companies and other organisations without proper authorisation from the countries and people concerned without compensatory payment Indian Parliament has recently cleared the second amendment of the Indian Patents Bill, that takes such issues into consideration, including ● Patent terms emergency provisions ● Research and development initiative.
  • 36. amrit_rj What is biopiracy ? ( 1 mark, 2016)
  • 37. amrit_rj What is biopiracy ? ( 1 mark, 2016) Biopiracy is the use of bioresources by multinational companies and other organization without proper authorization/compensation payment to the concern country /organisation.
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