12- BIOTECHNOLOGY AND ITS APPLICATION

CHAPTER NO.12 BIOTECHNOLOGY AND ITS APPLICATIONS

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WHAT IS TRANSGENIC ORGANISM?

A Transgenic organism is an animal, plant or microbe whose DNA is manipulated to possess and express an extra (foreign) gene by using genetic engineering techniques.

Gene that is artificially introduced into the genome of another organism is called a transgene.

The production of transgenic organisms using these transgenes is known as transgenesis .

A transgene will code for a protein, which corresponds to a particular trait.

Transgenes are introduced into the recipient organism's germ line. Because of the universality of genetic code, a transgene will allow transgenic organisms to produce the same protein as the original donor organisms.

The transfected organism gives birth to transgenic offspring which can further be bred to form a transgenic line.

Transgenic animals can be made by transfer of whole nuclei or transfer of whole individual chromosomes or fragments as well as by transfer of DNA.

Transgenic animals are also called transgenics or Genetically Modified

Organisms or GMOs.

Many bacteria, plants and animals are produced, which express foreign genes

inside them.

TRANSGENIC MICROORGANISMS:Transgenic bacteria produce many important substances for health and industry,because they reproduce quickly and are easy to grow.Bacterial cells can be genetically modified so that they have the gene for producing human insulin. As these modified bacteria grow, they produce human insulin. This

protein can be purified and supplied to diabetics GM bacteria are used in the production of enzymes such as milk-clotting enzymes for cheese production and food/feed additives such as aspartame and I-lysine.

TRANSGENIC PLANTS

Transgenic plants are plants that have been genetically engineered, using DNA

techniques to create plants with new characteristics.Today, approximately 90 percent of the corn, soybeans, and sugar beets on the market are GMOs. Genetically engineered crops produce higher yields, have a longer shelf life, are resistant to diseases and pests, and even taste better. These benefits are a plus for both farmers and consumers. For example, higher yields

and longer shelf life may lead to lower prices for consumers, and pest-resistant

crops means that farmers don't need to buy and use as many pesticides to grow

quality crops. GMO crops can thus be friendly to the environment than conventionally grown crops.

TRANSGENIC ANIMALS

A transgenic animal is one that has a foreign gene that has been inserted into its genome .

Transgenic rats, rabbits, sheeps, pigs , cows, and fish have been produced,although over 95 percent of all existing transgenic animals are mice

Transgenesis involves a number of methods like:

1. Transfer of the whole nucleus from a somatic cell into the enucleated egg of

the recipient animal.

2. Transfer of a part of dissected embryo into the enucleated unfertilized egg.

3. Transfer of a chromosome or chromosomal fragments

4. DNA microinjection technique. The new gene is introduced directly into the

fertilized ovum.

5. Gene targeting using embryonic stem cells: The gene is introduced into the

Embryonic stem cell soon after fertilization and then implanted into surrogate mothers.

1. TO STUDY NORMAL PHYSIOLOGY AND - DEVELOPMENT PARTICULARLY OF DIFFERENT GENES AND THEIR BIOLOGICAL EFFECTS

Transgenic animals can be specifically designed to allow the study of how

genes are regulated and how they affect the normal functioning of the body and

its development e.g., study of complex factors involved in growth such as an

insulin-like growth factor. 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.

2. STUDY OF DISEASE BY USING THEM AS MODELS:

Many transgenic animals are designed to increase the understanding of how

genes contribute to the development of diseases such as cancer, cystic fibrosis,

rheumatoid arthritis and Alzheimer. These are specially made to serve as models for human diseases, so that investigation of new treatments for diseases is made possible. Transgenic mice and rats are also being used as models for human diseases since they are easier to monitor, and have a

physiology corresponding to many human diseases..

3. TO STUDY VACCINE SAFETY TESTING:Transgenic mice are first to be used as laboratory animals in testing the safety of vaccines before they are used in humans .Transgenic mice are being used to

test safety of polio vaccine.

4. TO TEST CHEMICAL SAFETY:Transgenic animals are also used for the toxicity or safety testing procedures. If found reliable and successful they could replace the use of monkeys in order to test the safety of batches of the vaccine. Transgenic animals are made to carry the genes, which make them more sensitive to the toxic substances than the non-transgenic ones. They are then exposed to toxic substances and effects

are studied. The time required to obtain the results is less. This is known as

toxicity/safety testing.

6. TO OBTAIN INCREASED PRODUCTION OF BIOLOGICAL PRODUCTS:

i. In 1997, the first transgenic cow, Rosie, produced human protein enriched milk (2.4 grams per litre). The milk contained the human gene alpha-lactalbumin and was nutritionally a more balanced product for babies than that of natural cow milk.

ii. Transgenic sheep having different foreign genes have been produced.In the transgenic sheep, wool production was found more than non transgenic sheep.

iii. Sheep with the gene for growth hormone has also been formed , they yield more mutton.

iv. Transgenic fishes of many species like carp, catfish, salmon have been produced by microinjection of genes coding for human growth hormone.Transgenic fish are twice in size then than non transgenic fish.

7. INMOLECULAR FARMING:The method of manufacturing drugs in transgenic animals is called molecular farming. The transgenic animals can revolutionize the pharmaceutical industry by providing large quantities of valuabledrugs and proteins.

i) Transgenic goats have been used to produce tissue plasminogen activator which helps in dissolving blood clots and is useful for treating coronary

thrombosis.

ii)Female sheep having human anti haemophilic factor IX gene in their genome have been produced.

iii)Transgenic sheep having human protein (a-1-antitrypsin ) which is used to

treat emphysema.( using sheep), milk which can be used to treat coronary thrombosis .Thus by transgenesis the animals are genetically modified in such a way that they start acting bioreactors producing useful products in abundance and continuously.

8. ORGAN TRANSPLANTATION:Transgenic animals today are also being used in the field of organ transplantation.Pigs and mice are being used to grow organs like the heart, ear and pancreas

ISSUES RELATED TO TRANSGENIC TECHNOLOGY:

There may be health risks associated with transgenics.

There may be long term effects on the environment when transgenic animals are released into the field.

Various bioethics consider it is wrong to create animals that would suffer as a

result of genetic alterations.

CONCLUSION:Many transgenic animals have been successfully created for a variety of

purposes, and the prospects are enormous.

It holds great potential in many fields including agriculture, medicine and

industry. With proper research and careful use, transgenic animals can goa

long way in solving problems.

LET US KNOW WHAT WE HAVE LEARNT!

PART-A VERY SHORT ANSWER TYPE QUESTIONS:

A) MULTIPLE CHOICE QUESTIONS:

Q1. Maximum number of existing transgenic animals is of:

a) Cow

b) Mice

c) Fish

d) Pig.

Q2.Organisms that has had a foreign gene inserted into them is called:

a) Hermaphrodite

b) Transgenic Organisms

c) Transmuted Organisms

d) Transposon.

Q3.Today transgenic model exist for disease like:

a) Cold

b) Fatigue

c) Cystic fibrosis

d) Fever

Q4. Which of the following methods should be used to increase crop production without use of chemicals and fertilizers?

a) Use of ancient technique of agriculture

b) Use of genetically modified crops

c) Use of organic fertilizers

d) Use of more water

Q5. Gene used to treat emphysema is:

a) Trypsin 1

b) Beta -1 anti-trypsin

c) Alpha-1

d) Alpha -1 antitrypsin

B) STATE TRUE OR FALSE:

1. Transgenic mice are being developed for use in testing the safety of vaccines before they are used on humans.

2. Production of transgenic animals require transfections of eggs or embryos

3. The gene transferred to another organism artificially by technique of genetic

engineering is called wonder gene.

C) FILL IN THE BLANKS:

1. The first transgenic cow ,Rosie produced human milk with protein measured

to be

2. The protein secreted in the milk of transgenic goats can be used in treatment

of disease

A) MULTIPLE CHOICE QUESTIONS:

1. (b) mice (approximately 95% of transgenic animals are transgenic mice)

2 (b) Transgenic animals (Transgenic animals are those who have a foreign gene within them.)

3.(c) Cystic fibrosis (Transgenic animals serve as a model for human diseases. Cystic fibrosis is a genetic, hereditary disease that affects the lungs and digestive system of humans.)

4. (b) use of genetically modified crops.(Genetically modified crops may increase crop production many times without use of chemicals and fertilizers. )

5. (d) alpha- 1 anti-trypsin.(alpha- 1 antitrypsin is used to treat emphysema,

this disease causes breakdown of alveoli cells and reduces surface area for

exchange of gases.)

B) TRUE /FALSE:

1 True

2 True

3 False (The gene transferred to another organism artificially by technique of

genetic engineering is called transgene)

C) FILL IN THE BLANKS:

1. 2.4grams per litre

2. coronary thrombosis

PART B- SHORT ANSWER QUESTIONS:

Q1. What are transgenes and trans genesis?

Q2.The protein secreted in the milk of transgenic goats can be used in the

treatment of which disease?

Q3. Why transgenic mice are most preferred model of human diseases?

PART- C LONG ANSWER TYPE QUESTIONS:

Q1.What is transgenic animals? What are the categories of application of transgenic animals?

Q2. Write the role of transgenic animals in vaccine safety.

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INTRODUCTION

BIOTECHNOLOGY is the integration of natural science and organisms, cells,parts and molecular analogues for products and services useful to mankind.

BIOTECHNOLOGY HAS A WIDE RANGE APPLICATIONS:

a) Biopharmaceuticals

b) Therapeutics

c) Diagnostics

d) Genetically modified crops for agriculture

e) Processed food

f) Bioremediation

q) Waste treatment

h) Energy production

GREEN REVOLUTION succeeded in increasing the yield of crops by the use of agrochemicals but these are often too expensive so through genetic

engineering introduced GENETICALLY MODIFIED CROPS.

GENETICALLY MODIFIED PLANTS OR TRANSGENIC PLANTS:The plants in which foreign genes have been introduced through genetically engineering are called TRANSGENIC PLANTS.

METHOD FOR MAKING OF TRANSGENIC PLANTS:

Isolate DNA that codes for the protein you want to express.

Insert the DNA into a plasmid.

Insert the plasmid into bacteria. Grow a large amount of bacteria containing this plasmid.

Dip the flowering plant into a large amount of bacteria.

Give bacteria the opportunity to insert the DNA into the plant cells.

Select for plants that have the insertion.

INSECT RESISTANCE IN TRANSGENIC PLANTS:

Soil bacterium Bt. Cotton (Bacillus thurengenesis) produces proteins that

kill insects like lepidopteron (bud worm), coleopteran’s (beetles) and dipterans (flies). These crystals contain a toxic insecticidal protein. The Bt.

toxin form proteins exist as inactive PROTOXINS but once insects ingests

the inactive toxin it is converted into an active form of toxin that solubilizes

the crystals. The activated toxins bind to the surface of midgut epithelial

cells and create pores which cause cell swelling and lysis and finally cause

death of the insect.

Bt. toxins genes were isolated from Bacillus thurengenesis and incorporated into crop plants such as cotton.

Most of Bt. toxins are insect-group specific. The toxin is coded by a gene named Cry. For e.g. proteins encoded by the genes crylAc and

cryll[Ab control the cotton bollworms. Similarly, cry[Ab has been introduced in Bt.corn to protect the same from corn borer.

PEST RESISTANCE IN TRANSGENIC PLANTS: (PROTECTION AGAINST NEMATODES):

A nematode Meloidogyne incognitia infects the roots _of tobacco plants causing a reduction in yield. It can be prevented by RNA interference (RNAi)strategy. RNAi is a method of cellular defence in all eukaryotic organisms.

HERBICIDE RESISTANT TRANSGENIC PLANTS:

Weeds such as Striga decrease crop yields and quality by competing with crop plants for light, water and nutrients. Weeds are to be removed with the help of herbicides. For e.g. ROUNDUP READY transgenic plant has been

produced and commercialised.it is tolerant to herbicide ROUNDUP (trade name).

MOLECULAR FARMING:Production of proteins encoded by transgenes in animals/crops; the

proteins recovered from milk, urine, seeds is called MOLECULAR FARMING.

ADVANTAGES OF TRANSGENIC PLANTS:

Pest resistant crops

Tolerance against environment stress

Reduction in post- harvest losses

Herbicide resistance

Increasing nutritional value of food

DISADANTAGES OF TRANSGENIC PLANTS:

Environmental hazards

Human health risks like allergies

PART: A_ VERY SHORT ANSWER TYPE QUESTIONS:

(A) MULTIPLE CHOICE QUESTIONS:

1. Bt cotton variety that was developed by the introduction of toxin gene of

Bacillus thurengenesis (Bt) is resistant to:

(a) Insect pests

(b) fungal diseases

(c)plantnematodes

(d) insect predators

2. Consumption of which one of the following foods can prevent the kind of blindness associated with vitamin ‘A’ deficiency?

(a) ‘Flavr Savr tomato

(b) Canolla

(d) Bt-Brinjal

3. The process of RNA interference (RNAi) has been used in the development of plants resistant to:

(a) Nematodes

(b) fungi

(d) insects

4. Golden rice is a transgenic crop of the future with the following improved trait-

(a) Insect resistance

(b) high lysine (essential amino acid) content

(d) high vitamin-A content.

5. In RNAi, the genes are silenced using-

(a)ds-RNA

(b)ss-DNA

(c)ss-RNA

(d)ds-DNA

B. TRUE AND FALSE:

1. Meloidegyne incognitia infect stem of tobacco plant

2. RNA interference is essential for the cell defense.

C. FILLIN THE BLANKS:

1. The alkaline pH in the midgut of insect larvae triggers the activation of .

2. Using Vectors, nematode-specific genes are introduced into host plants.

3. The genes crylAb and cryllAb produce toxins against and ,respectively.

ANSWERS KEY: PART -A

A. MULTIPLE CHOICE QUESTIONS:

1. (a) Insect pests

2. (c) Golden rice

3. (a) Nematodes

4. (d) High vitamin-A content

5. (a) ds-RNA

B. TRUE OR FALSE:

1. FALSE, It infects roots of tobacco plant.

2. TRUE

C. FILL IN THE BLANKS:

1. Bt.toxin

2. Agrobacterium

3. Corn borer, cotton bollworm.

PART: B SHORT ANSWER TYPE QUESTIONS:

1. What are Cry proteins? Name an organism that produces it.

2. What are the problems associated with GMF.

3. Nematode resistant transgenic plants have been produced. Explain diagrammatically.

PART: C LONG ANSWER TYPE QUESTIONS:

1. What is the importance of Biotechnology in agriculture explain in detail.

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INTRODUCTION

Recombinant DNA Technology in Biotechnology plays a very important role in large

scale productions of safe and more effective recombinant therapeutics.

RECOMBINANT THERAPEUTICS-Recombinant pharmaceuticals or therapeutics is created by inserting genes from one species into a host species, often yeast or bacteria.For example, In case of insulin production, genes coding for human insulin are inserted into bacteria. Bacteria produce insulin, which is harvested and used for diabetic patients.

ADVANTAGES OF RECOMBINANT THERAPEUTICS:

Recombinant therapeutics never induce unwanted immunological responses as is

common in case of similar products isolated from non-human resources or other

animals which contain other proteins and generate immune responses.Recombinant therapeutics is produced in very large amounts and also very easy to produce.

These drugs are very effective with no any kind of side effects.Production of these drugs are time and money saving.30 recombinant therapeutics have been approved for human use on word level but in India 12 of these are approved and presently being marketed.

NATURAL PRODUCTION OF INSULIN:Diabetes- It is a type of disease when blood glucose is too high.Insulin- This is a hormone produced by pancreas. It converts extra blood glucose

into glycogen which further stored in liver.

In humans insulin is synthesized as a prohormone (inactive form) and consists of

three polypeptide chains: chain A, chain B and an extra chain called C-peptide.

During its processing C-peptide is removed and chain A and chain B joined by

disulfide bonds. Then hormones become fully mature and functional.

In diabetic patient enough insulin not produced which results in high blood sugar level.In past insulin isolated from pancreas of slaughters animal pig and cow. Sometimes it cause allergy.

BIOTECHNOLOGICAL PRODUCTION OF INSULIN:

In 1983 Eli Lilly an American company took initiative. Company prepared two DNA

sequence correspond to A and B chain of insulin and introduce them into plasmid

of Escherichia coli bacteria separately and produce corresponding A and B chain

separately in separate culture medium or in separate bioreactor.In next step chain A and B extracted and combined by creating disulfide bond to form final product (Human insulin) or HUMULIN.

LET US KNOW WHAT WE HAVE LEARNT!

PART: A VERY SHORT ANSWER TYPE QUESTIONS:

(A) MCQs:

1. Insulin hormone consist of of —

a. Polypeptide chain

b. Lipid molecules

c. Oligosaccharides

d. Nitrogen oxides molecules

2. Recombinant pharmaceutical insulin produced by using-

a. Fungi

b. Eukaryotic cell

c. Bacteria

d. Virus

3. Which bond hold together amino acids chain in insulin hormone?

a. Hydrogen bond

b. Peptide bond

c. Disulfide bond

d. Ester bond

4. How many recombinant pharmaceutical product approved in India for

human use?

a. 30

b. 12

c. 22

d. 15

5. Which gland produces insulin hormone?

a. Pituitary gland

b. Thyroid gland

c. Pancreas gland

d. Liver

(B) TRUE/FALSE:

1. Insulin hormone consists of lipids and organic acid.

2. Insulin isolated from other slaughtered animal not cause any kind of

allergy in human being.

3. In 1983 Eli Lilly an American company took initiative to produce

recombinant insulin.

(C) FILL UPS:

1. ____ Number of recombinant pharmaceutical products approved for

human use on word level.

2. It is very easy to produce on a large scale with the help of

biotechnology.

ANSWER KEY: PART -A

MCQs:

1. a Polypeptide chain

2. c Bacteria

3. c Disulfide bond

4. b12

5. c Pancreas gland

TRUE/FALSE:

1. false

2. false

3. true

FILL UPS:

1. 30

2. Recombinant therapeutics

PART:B SHORT ANSWER TYPE QUESTIONS:

1. How is Prohormone different from hormones?

2. Why is recombinant therapeutics safe for human use?

3. What is c-peptide?

PART: C LONG ANSWER TYPE QUESTIONS:

1. Describe the role of biotechnology in large scale production of recombinant pharmaceuticals with the help of an example insulin.

A136

INTRODUCTION

GENE THERAPY is a collection of methods that allows correction of a gene defect that has been diagnosed in a child/embryo.

Genes are inserted into a person’s cells to treat a disease.

Correction of a genetic defect involves delivery of a normal gene into the

individual or embryo to take over the function and compensate for the non-

functional gene.

The first clinical gene therapy was given in 1990 to a 4-year old girl with

Adenosine De Aminase (ADA) deficiency.

GENE THERAPY is therapeutic treatment of defective heredity by introduction of normal healthy and functional genes to silence the defective genes.

ADA DEFICIENCY:ADA (adenosine deaminase) enzyme is crucial for the immune system to

function.

In some children ADA deficiency can be cured by bone marrow transplantation or by enzyme replacement therapy.

STEPS OF GENE THERAPY:

Lymphocytes from the blood of the patient are grown in a culture outside the body.

A Functional ADA cDNA (Complementary DNA produced by reverse transcription using a retroviral vector) is introduced into these

lymphocytes which are returned to the patient.

Now these cells are not immortals, so the patient requires periodic infusions of such genetically engineered lymphocytes.

For a permanent cure, the genes are isolated from cells producing ADA & are introduced into cells at early embryonic stage. This is

however not possible.

Therefore in 1990 midway approach is adopted to cure a 4-year old girl,Ashanti.

MAJOR REQUIREMENT OF GENE THERAPY USING RETROVIRUSIS:

The isolation of the desired gene

Amplification of the desired gene

Aretrovirus for removal of viral genes

Insertion of desired gene

Helper retrovirus lacking packing sequence.

PROCEDURE:

ADA gene was taken out from leucocytes of a healthy person.

It is amplified in bacteria and inserted in retrovirus, from which viral!genes had been taken out.

A helper retrovirus devoid of packing sequence was also prepared.

Lymphocyte stem cells from bone marrow of patient were taken out.

These lymphocytes are now injected with two types of retroviruses,one carrying ADA gene and other helper.

The normal ADA gene begins to express in the lymphocyte stem cells, now called transformed stem cells.

The transformed healthy stem cells, now introduced into the bone marrow of the patient.

The immune system of the patient now becomes functional.

LET US KNOW WHAT WE HAVE LEARNT!!

PART: A VERY SHORT ANSWER TYPE QUESTIONS:

A) MULTIPLE TYPE QUESTIONS:

Q1: A genetic disorder can be cured through:

a) rDNA technology

b) embryo transfer

c) gene therapy

d) all of the above

Q2: Gene therapy in humans was first practiced to cure:

a) Cystic fibrosis

b) Haemophilia

c) Thalassemia

d) ADA deficiency/severe combined immuno deficiency disease

Q3: In gene therapy the genetic defect is corrected by delivery of which gene?

a) Incorrect

b) Mutant

c) Normal

d) Jumping

Q4: ADA stands for:

a) Adenosine nucleotide amine

b) Vitamin A deficiency

c) Adenosine deaminase

d) A double bond amine

Q5: ADA deficiency can be permanently cured by:

a) Bone marrow transplantation

b) Enzyme replacement therapy

c) Gene therapy at early embryonic stages

d) All of these

B) TRUE/FALSE:

1) Lymphocytes are immortal.

2) ADA enzyme is crucial for immune system to function.

3) A functional ADA cDNA is introduced using retroviral vector.

C) FILL UPS:

1) The first clinical gene therapy was givenin tt.

2) The patient requires periodic infusionof_ tt.

ANSWER KEY: PART-A

A) MULTIPLE TYPE QUESTIONS:

1. C) Gene Therapy

2. D) ADA deficiency/severe combined immuno deficiency disease

3. C) Normal

4. C) Adenosine deaminase

5. C) Gene therapy at early embryonic stages

B) TRUE/FALSE:

1) False

2) True

3) True

C) FILL UPS:

1) 1990

2) Genetically engineered lymphocytes.

PART: B SHORT ANSWER TYPE QUESTIONS:

Q1: What is meant by gene therapy?

Q2: What is ADA Deficiency?

Q3: How ADA Deficiency can be cured?

Q4: What is retroviral vector?

PART: C LONG ANSWER TYPE QUESTIONS:

Q1: Illustrate in detail gene therapy using ADA deficiency?

A137

INTRODUCTION

ETHICAL ISSUES:Certain acts are regulated by communities to consider their legitimacy.

Such regulations are called ETHICS.ETHICAL ISSUES occur when a given decision, scenario or activity creates a conflict with a society’s moral principles.

ETHICAL ISSUES:

I. Introduction of a transgene from one species into another species violates the integrity of species.

ll. Biotechnology may lead to unforeseen risks to the environment,including risk to biodiversity. It can disturb the existing ecological balance.

Ill. Utilization of animals !In biotechnology causes great suffering to them.

IV. There is another biological damage .it can accidentally create new infectious agents.

V. When animals are used for production of pharmaceutical proteins they are virtually reduced to the status of a factory.

BIO PIRACY:Pirates in general terms were blood thirsty. They stole and killed other to enrich themselves. The bio pirates are slightly different. They do not kill, they patent. They are completely protected by law asthe result, no body hurts them.

EXAMPLES OF BIOPIRACY:For thousands of years Neem is being used in India for killing pests and as medicine. One of the American companies patented Neem.As a result anybody using Neem will have to pay for it. This was called legalized theft.The free heritage of seeds or knowledge can be protected by patent. The basic meaning of PATENT is intellectual property right or IPRs. If the product is not patented, it may be freely shared between everybody who required them, but after getting its patent one will have to buy it to use it legally.

BIOPATENT help in economic growth for individual involved and country concerned. Biopatents are being done due to their ethical and political values also. Sometimes the unplanned Genetic modification of organisms may produce unpredictable harmful results. So the Indian government set up

genetic engineering approval committee (GEAC), which keeps an eye on the validity of GM approach and the safety of GM organisms for the public services like food and medicine services.

A nice example of bio patency of indigenous biological resources by an influential corporate is that Basmati rice an important food grains crop being grown in the Asian region even sent thousands of years in 1977.

US patent and trade mark office granted its bio patent to an American company though it was produced by crossing Indian basmati rice variety with semi-dwarf variety.

BIOETHICS:Certain acts are regulated by communities to consider their legitimacy. Such regulation is called ethics. Bioethics includes standards followed by us to

regulate the activities linked with biological activities.BIOETHICS concerns related to biotechnology are:

I. For the production of pharmaceutical products like Protein many animals are

reduced to thestatus of factory.

ll. Animals undergo great physical sufferings while performing experiments on them.

lll. There is violation of integrity of species due to transferring of a transgene

from one speciesto another.

IV. Concept of humanness is diluted due to introduction of human genes to various animals orvice versa.

V. Biotechnological studies have been considered as mere exploitation.Biodiversity may be affected due to biotechnological experiments, thus posing a threat to environment.

BIOPATENT:Government grants the patent to certain inventors, a permission to use invention for commercial purpose. Patent is given for:

a) Producing new product or invention.

b) Modified and improved earlier invention.

c) Technical know-how.

d) New designing concept.

BIOPATENT are being granted by the government for biological entities and products derivedfrom them.BIOPATENT are given:

a) For discovery of new strains of microorganisms.

b) Transgenic (genetically modified) pants and animals.

c) Discovery of cell lines.

d) New biotechnological methods.

e) Unearthing the DNA sequences.

f) Formation of new product.

g) New product techniques.

Some Interesting Aspects of Bio Piracy:

Mae-wan ho biologist from Open University, UK (1999) is of the view that transgenic agriculture is needed to feed the world .The tightening of corporate monopoly on food by patenting the seeds may cause famine.

It also diverts the various countries to successful Implementation of sustainable, organic agriculture which can ensure food security and better health for ail.

Adalberto Antonio, judge of state of Amazons points out that Harry Wickham took about 70,000 seedson behalf of new gardens, Britain. This only act plunged the state of Amazons in to starvation for year’s altogether.

American countries are also opposing the new patents on life and to commercialization of science.

Several scientists have called for various governments to improve a moratorium on global releases oftransgenic crops and to ban patents on living organisms.

LET US KNOW WHAT WE HAVE LEARNT!!

PART: A VERY SHORT ANSWER TYPE QUESTIONS:

A) Multiple choice questions:

1. bt cotton is not :

a) AGM plant

b) Insect resistant

c) Abacterial gene expressing system

d) Resistant to all pesticides

2. GEAC stands for:

a) Genome engineering action committee

b) Ground Environment Action committee

c) Genetic engineering approval committee

d) Genetic and environment approval committee

3. Golden rice is:

a) A variety of rice grown along the yellow river in china

b) Long stored rice having gene for b-carotene

c) Wild variety of rice with yellow colored grains.

d) All the above

B) Fill in the blanks:

1. Plants , bacteria, fungi and animals whose genes have been altered by

manipulation called

2. In GM plants, genetic modification enhances

3. Basmati rice is distinct for its unique aroma and

C) True/ False:

1. Genetic engineering approval committee is expansion of GEAC.

2. First Genetically modified plant commercially releases in India is Bt-cotton..

3. Golden rice is rich in vitamin A.

ANSWER KEY: PART -A

A) Multiple choice questions:

1. d) Resistant to all pesticides

2. c) Genetic engineering approval committee

3. c) Wild variety of rice with yellow colored grains.

B) Fill in the blanks:

1. genetically modified organisms

2. nutritional

3. flavour

C) True/ False:

1. True

2. True

3. True

PART: B SHORT ANSWER TYPE QUESTIONS:

1. Right a short note on Bio piracy.

2. What are transgenic organisms?

3. What are disadvantages of GM crops?

PART: C LONG ANSWER TYPE QUESTIONS:

1. What do you mean by bio-patent? Write its uses and abuses?

A138

RECAPITULATION

Biotechnology has given to humans several useful products by using microbes, plant, animals and their metabolic machinery.

Genetic engineering involves the isolation of specific genes, making of copies of genes and their transfer to target genomes.

Genetically Modified organisms have been created by using methods other than natural methods to transfer one or more genes from one organism to another, generally using techniques such as recombinant DNA technology.

Bacteria, yeast and cultured plant and animal cells are commonly used as hosts for recombinant DNA experiments.

Agriculture has been a major focus of genetic engineering activity.

GM plants have been useful in:

1. Increasing crop yields

2. Reduce pest harvest losses

3. Make crops more tolerant of stresses

4. Improve nutritional value of foods

5. Produce pest resistant crops

Transgenic animals are also used to understand how genes contribute to the

development of a disease.

In gene therapy, a faulty gene is replaced with a normal healthy gene, to treat disease where other medical approaches are not effective like cancer,AIDS and Haemophilia.

Despite benefits, the ethical, social and legal implications of these potent

gene technologies have led to considerable public concem over the possibility of accidentally producing new pathogens or “Genetic Monsters”

Q1. Crystals of Bt toxin produced by some bacteria do not kill the bacteria themselves because:

a) bacteria are resistant to the toxin

b) toxin is immature;

c) toxin is inactive;

d) Bacteria encloses toxin in a special sac.

Ans.1 (C) toxin is inactive: In bacteria, the toxin is present in an inactive form, called protoxin, which gets converted into active form when it enters the body of an insect.

Q2. What are transgenic bacteria? Illustrate using any one example.

Ans.2. Transgenic bacteria contain foreign gene that is intentionally introduced into its genome. They are manipulated to express the desirable gene for the production of various commercially important products.

An example of transgenic bacteria is E.coli. In the plasmid of E.coli, the two DNA sequences corresponding to A and B chain of human insulin are inserted, so as to produce the respective human insulin chains. Hence, after the insertion of insulin gene into the bacterium, it becomes transgenic and

starts producing chains of human insulin .Later on, these chains are extracted from E. coli and combined to form human insulin.

Q3. Compare and contrast the advantages and disadvantages of production of genetically modified crops.

Ans. 3

Q. 4) What are cry proteins? Name an organism that produces it. How has man exploited this protein to his benefit?

Ans.4 1. Cry proteins refer to the protein crystals containing a toxic insecticide.

2. It is produced by soil bacterium, Bacillus thuringiensis.

3. The genes encoding cry proteins called Bt toxic genes were isolated from

B.thuringiensis and incorporated several crop plants such as Bt cotton, Bt corn etc., to provide resistance against insect pests .

Q.5) What is gene therapy? Illustrate using the example of adenosine deaminase (ADA) deficiency?

Ans.5 Gene therapy is a process of introduction of DNA into an organism in

order to treat a disease. It is used to replace a missing gene product or to

correct mutant alleles. e.g. - Human beings.

ADA (adenosine deaminase deficiency) is a rare genetic disorder caused due to deletion of the gene for adenosine deaminase. This enzyme is critical for the normal functioning of the immune system. This disorder can be

treated by gene therapy wherein the gene is transferred into early embryonic cells of the bone marrow for permanent utilization.

Q.6) Diagrammatically represent the experimental steps in cloning and

expressing a human gene ( say the gene for growth hormone ) into a bacterium like E. coli?

Ans.6 DNA cloning is a method of producing multiple identical copies of specific template DNA. It involves the use of a vector to carry the specific foreign DNA fragment into the host cell. The mechanism of cloning and transfer of gene for growth hormone into E. Coli is represented below:-

Q.7) Can you suggest a method to remove oil (hydrocarbon) from seeds based on your understanding of r DNA technology and chemistry of oil?

Ans.7 Recombinant DNA technology or rDNA is a technique which is used for the manipulation of the genetic material of an entity in order to obtain desired results. The genes for the formation of oil in the seed should be

identified. The appropriate genes should be removed with the help of restriction endonucleases. Such DNA should then be treated with DNA ligases to make seal DNA at the broken ends. These cells when grown

aseptically on nutrient medium will differentiate into a new plant whose seeds

will not have oil in them.

Q.8) Find out from internet what is golden rice.

Ans. 8 Golden Rice, a variety of rice, Oryza sativa is a genetically modified crop, which is developed as a fortified food to supply them to areas where there is scarcity of dietary vitamin A, as Golden Rice is richly supplied with vitamin A.Golden Rice consists of a precursor of pro vitamin A, known as the beta carotene, inserted into the rice through the process of genetic engineering.This is a simpler and low priced alternative. It was developed at Swiss Federal Institute of Technology by professor Ingo potrykus and Peter Beyer.

Q.9) Does our blood have proteases and nucleases?

Ans.9 No, blood does not have proteases and nucleases. But some proteases do exist in its inactive form. If it would have been found in blood and cells, it would have been digested.

Q.10) Consult internet and find out how to make orally active protein pharmaceutical. What is the major problem to be encountered?

Ans.10 Orally active protein pharmaceuticals contain biologically active material such as proteins, antibodies and polymeric beads. It is administrated orally into the body through various formulations.These proteins are used for treatment of various diseases such as Hepatitis B, Herpes, and Influenza etc. and are also used as vaccines.However the oral administration of these proteins has some problems related to it. Once these proteins are ingested, the proteases present in the stomach juices denature the protein. As a result, there effect will be nullified.

Hence it is necessary to protect the therapeutic protein from digestive enzymes, if taken orally. This is the reason for the proteins to be injected directly into the target site.

A139

RECAPITULATION

Dear students we have discussed the chapter no. 12 “Biotechnology and its applications” and its NCERT questions in previous daily dose assignments. Now let us revise important concepts of this chapter.

Biotechnology is the process of using living organisms or their enzymes or their molecules to produce products and processes for the welfare of mankind.

Application of DNA manipulation ranges from cloning genes to cloning organisms including transgenic microbes, agriculturally important crops and farm animals. Humans have used biotechnology in the improvement of the

quality of life, in the field of health and food production.

APPLICATIONS IN MEDICINES:

1. GENETICALLLY ENGINEERED INSULIN: -Insulin is produced in 8 cells of islets of Langerhans synthesised as proinsulin which is inactive form of insulin. Insulin is made up of two short polypeptide chains Chain A (having 21 amino acids) and Chain B (having

30 amino acids) linked by disulphide bonds. But proinsulin contains one more polypeptide chain called C chain which acts as linker chain, helps to bind sulphur molecules on both A and B chain. As soon as disulphide

bond is formed C chain disappear and active insulin molecule is formed.

Earlier insulin used by diabetics were extracted out of pancreas of slaughtered animals. In some patients, due to insulin received by animals,other types of reaction against allergies or external proteins could be started.

To avoid these situations, in 1983, a US company named Eli Lily designed two DNA sequences coding for A chain and B chain and

introduced them into the plasmid of E. Coli. Chains A and B were produced separately extracted and combined by creating disulphide

bonds of form human insulins.

Insulin (Humulin) was the first hormone which is produced artificially by culturing bacteria by employing recombinant DNA technology.

GENE THERAPY:Gene therapy in humans is to replace “a faulty gene” by a normal

healthy functional gene.

The first clinical gene therapy was given in 1990 to a 4-year-old girl with Adenosine De Aminase (ADA) deficiency. The enzyme is crucial for the immune system to function.

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 (using a retroviral vector) is then introduced into these lymphocytes which were then returned to patient. But life of lymphocytes is short so such patients are

required to give infusion at the short intervals regularly.

However, if this treatment is done at early embryonic stage, is could be a permanent cure.

MOLECULAR DIAGNOSIS:DNA technology has provided a broad range of tools to diagnose

various diseases like food poisoning Salmonella, pus forming Staphylococcus, hepatitis virus, HIV and so on.

By testing the DNA of prospective genetic disorder in carrier parents, their

genotype can be determined and their chances of producing an afflicted child can be predicted.

ELISA, PCR are some of the techniques by which early detection of disease

can be done. ELISA (Enzyme Linked Immuno-Sorbent Assay) is based on the principle of antigen antibody interaction.

In suspected AIDS patients, PCR for the identification of HIV is being used routinely now a day.

It is also used to detect the mutations occurring in the genes of suspected

cancer patients.

TB can also be detected by the antigen antibody reaction.

APPLICATIONS IN AGRICULTURE:Though Green Revolution succeeded in tripling the food supply but it is not possible to fill the growing needs of human population. For this, use of Genetically Modified Crops is the only solution.

GENETICALLY MODIFIED ORGANISMS:Plants, bacteria, fungi and animals, whose genes have been converted by manipulation, are called Genetically Modified Organisms.

The use of GM plants is beneficial in many ways:

i. GM crops are more tolerant to abiotic stresses (cold, drought, salinity,heat).

ii. Less dependency on chemical pesticides.

iii. Helped to reduce post-harvest losses.

iv. Increased efficiency of mineral usage by plants.

v. Increase in nutritional level of food e.g., Vitamin A enriched rice.

Vi In the use of biotechnology in agriculture it is the production of pest resistant crops that reduced the use in the amount of pesticide.

Bacillus thuringiensis (Bt) toxin genes are cloned by bacteria and introduced in plants to produce pest resistant crops e.g., Bt cotton, Bt rice, Bt tomatoes, Bt potatoes and Bt soyabean etc.

A. Bt cotton: - Some strains of Bacillus thuringiensis produce proteins that can kill certain insects. But these proteins are active only when they come in contact with alkaline pH. As plants do not have alkaline pH, the specific protein remains inactive. But when any insect or larva feed on

such plants, it will die.

B. PEST RESISTANT PLANTS: - To obtain pest resistant plants by RNAi (RNA interference techniques) e.g., the nematode Meloidegyme

incognitia reduces the yield of tobacco plant.

In tobacco plant dsRNA is formed by using Nematode gene which acts as defence mechanism.

When nematode enters the plant or feed on such plant, MRNA formed by its own gene binds with dsRNA formed from same gene in

the plant. As they bind mRNA cannot produce proteins required for the growth of nematode.

C. OBTAINING HIRUDIN from the seeds of Brassica napus by introducing hirudin producing gene in the plant.

D. FLAVR SAVR TOMATOES : -Complimentary DNA of enzyme Poly Galact Uronase is introduced in plant which silenced the production

of Pectin Protein and delayed the ripening of fruit.

E. GOLDEN RICE is the transgenic variety of basmati rice which gives high yield and rich in Vitamin A. It is used to cure the deficiency of Vitamin A.

TRANSGENIC ANIMALS:Animals that had their DNA manipulated to possess and express an extra gene are called transgenic animals. Transgenic rats, rabbits,pigs, sheep, cows and fish have been produced.

USES OF TRANSGENIC ANIMALS:

1. To study normal physiology and development controlled by various genes.

2. To study how genes contribute to the development of diseases.

3. Transgenic that produce useful biological products can be created by introduction of DNA (gene) which codes for a particular product.

In 1997, the first transgenic cow Rosie produced human protein enriched milk, which was more beneficial for human infants than

ordinary cow milk.

4. Transgenic mice are developed for use in testing the safety of vaccines before they are used on humans.

5. Transgenic help in chemical safety testing.

ETHICAL ISSUES:Genetic modification of organisms can have unpredictable results when

such organisms are introduced into the ecosystem.

GEAC (Genetic Engineering Approval Committee):set up by Indian Government make decision regarding the validity and safety of GM research.

BIOPIRACY

BIOPIRACY: is the term related to using biological resources without proper authorisation and compensatory payments from multinational companies and other organisations to a nation or its people.

The Indian Parliament has recently cleared the second amendment of the Indian Patients Bill that takes such issues into consideration.

LET US KNOW WHAT WE HAVE LEARNT!!

(A) MULTIPLE CHOICE QUESTIONS:

1. Insulin consists of two short polypeptide chains; chain A and chain B, that are linked together by:

a.) Hydrogen bridges

b.) Ester bridges

c.) Peptide bridges

d.) Disulphide bridges

2. The Bt toxin is produced by a bacterium called:

a.) Bacillus termegiensis

b.) Bacillus thermogiensis

c.) Bacillus thuringiensis

d.) Bacillus trichonegiensis

3. Golden colour of Rice is due to occurrence of:

a.)Vitamin A

b.)Vitamin C

c.) Vitamin K

d.)Vitamin Be

4. Transgenic animals are used in:

a.)Normal physiology and development

b.)Study of disease

c.) Vaccine & Chemical safety testing

d.)All of the above.

5. In RNAi, genes are silenced using:

a.)ss RNA

b.)ds DNA

c.)ds RNA

d.)ss DNA

(B) FILL IN THE BLANKS:

1. Flavr savr is the variety of tomato.

2. Transgenic hirudin is obtained from .

3. was the first hormone which is produced

artificially by using recombinant DNA technology.

4. A transgenic plant having high storage protein is .

5. part of tobacco plant is infected by incognita.

(C) TRUE/ FALSE:

1. SCID is caused by the defect in the gene for the enzyme Adenosine deaminase.

2. The growth of nematodes is controlled by using RNA interference technique.

3. Bt toxin in coded by a gene named Sad.

4. ELISA is the test for HIV.

5. Salt tolerant transgenic has been developed for tomato.

ANSWER KEY: PART - A

1.) (d) Disulphide bridges

2.) (c) Bacillus thuringiensis

3.) (b) Vitamin C

4.) (d) All of the above

5.) (c) ds RNA

1.)transgenic

2.)Brassica napus

3.)Insulin (Humulin)

4.)Potato

5.)Root

1.)True

2.) True

3.)False: Bt toxin is coded by a gene named cry.

4.)True

5.) True

PART B: SHORT ANSWER TYPE QUESTIONS:

1. What is GMO? How does it differ from a hybrid?

2. What is ELISA?

3. What is Biopiracy?

4. What is GEAC? What are its main objectives?

5. How are ‘cry’ and ‘CRY’ different from each other?

PART C: LONG ANSWER TYPE QUESTIONS:

1. Explain five areas where biotechnology has influenced human lives?

2. What are transgenic animals? Enlist any four areas where they can be

used?

A216

RECAPITULATION

The use of biology to develop technologies and products for the welfare of human being is known as biotechnology.e Biotechnology has a wide spread applications in the welfare of human beings.

Biotechnology Application in Agriculture: Biotechnology has different application in agriculture. It can be used in agro-chemicals, organic agriculture, and genetically engineered crop-based agriculture. It can be used to produce genetically modified organisms. Genetically modified organisms can be obtained by alternation in their genetic materials.

Genetic medication has: Production of crops which are resistant to abiotic factors such as heat, cold etc., Pest-resistant crops, Help to reduce post-harvest losses, Minerals can be used efficiently by the plants, Food with enhanced nutritional values, eg. Golden rice i.e. Vitamin ‘A’.

The most important application in the field of biotechnology is the formation of pest resistant plants.

Bt Cotton: Bt toxin is produced by a bacterium called Bacillus thuringiensis. Bt toxin gene has been cloned from the bacteria and been expressed in plants to provide resistance to insects without the need for

insecticides. The toxin is coded by a gene crylAc name cry. There are a number of them, for example, the proteins encoded by the genes crylAc and cryllAb control the cotton bollworms.

Another Pest Resistant Plants: Various pests affect the plants which causes loss as well as decrease in the yield of the plants. A nematode meloidegyme incognitia infects the roots of tobacco plants and cuases s

decrease in the yield of the plants. To prevent this, RNA interference technology was used.

Biotechnology Applications in Medicine: Biotechnology has contributed a lot in medicine industry. Tne use of biotechnology in medicine is known as medicinal biotechnology. They provide methods for

the formation of genetically modified insulin known as humulin. This helps in treatment of large number of diabetes patients.

Gene Therapy: To correct the heredity disease, gene therapy is used.The correction of gene defects is known as gene therapy. A normal gene is inserted into the individual or in an embryo to replace the abnormal

gene.

Molecular Diagnosis: It helps in disease diagnosis by various techniques such as ELISA, PCR and recombinant DNA technology.

Ethical Issues: GEAC: Genetic engineering Approval Committee is a committee set of by the Indian Government of oversee all decisions regarding GM research and the safely of GMOS for public use.

Biopiracy: Use of bio-resources by commercial and multinational companies and other organizations without the appropriate authorization and permission from the countries and people concerned without making

the compensatory payment.

1 MARKS Que/Ans (MCQ AND DEFINITION TYPE)

Q. 1 Which of the following technique can serve the purpose of early diagnosis of a disease: (a) Recombinant DNA technology

(b) PCR

(d) All of these

Ans.(d) All of these

Q.2 First hormone produced artificially by culturing bacteria is:

(a) Insulin

(b) Thyroxine

(d) Adrenaline

Ans.(a) Insulin

Q.3 A transgenic animal has:

(a) Foreign DNA in its cells

(b) Foreign DNA in some of its cells

(d) Both B and C

Ans.(a) Foreign DNA in its cells

Q.4 Which of the following peptide chain is not present in mature insulin:

(a)A

(b)B

(c)C

(d) None of these

Ans.(c)C

Q.5 ELISA is used to detect viruses where: (a) DNA probes are required

(b) Southern blotting

(d)Catalase is the key reagent

Ans.(c) Alkaline phosphatase is key reagent

Q.6 Golden rice is a transgenic crop of the future with the following improved trait:

(a) High lysine (essential amino acid) content

(b) Insect resistance

(d) High vitamin A content

Ans.(d) High vitamin A content

Q. 7 Bt cotton is not:

(a) a GM plant

(b) Insect resistant

(d) Resistant to pesticides

Ans.(d) Resistant to pesticides

Q.8 Antitrypsin is:

(a) An antacid

(b) An enzyme

(d) Use to treat emphysema

Ans.(d) Use to treat emphysema

Q.9 What is biotechnology?

Ans.The use of biology to develop technologies and products for the welfare of

human being is known as biotechnology.

Q.10 What is transgenic bacteria?

Ans.Those bacteria whose genes are treated by genetic engineering, are called transgenic bacteria e.g. E. coli bacteria having human insulin gene.

Q.11 Write a short note on biopiracy.

Ans.It is the use of bioresources, such as genetically engineered viruses,bacteria, plants and animals by multinational companies and _ other organisations without proper authorisation from concerned authorities and

people.

Q.12 Why is proinsulin so called? How is insulin different from its?

Ans.It is the prohormone which needs to be processed before it becomes a fully

mature and functional hormone/insulin. Proinsulin has three polypeptide chains (A, B and C), whereas insulin has only two (A and B) polypeptide chains.

Q.13 What is gene therapy? Name the first clinical case where it was used.

Ans.Gene therapy is a collection of methods that allows the correction of a defective gene. The first clinical gene therapy for adenosine deaminase (ADA) deficiency.

Q.14 What are cry genes? In which organism are they present?

Ans.1. The genes which code for the Bt toxin proteins, are called cry genes.

2. They are present in the bacterium,Bacillus thuringiensis.

Q.15 What is Ti-plasmid?

Ans.The Ti-plasmid, which stands for tumor inducing, is a piece of DNA that

occurs in soil inhabiting plant pathogenic bacterium.

Q.16 What are transgenic plants?

Ans.These plants whose genes have been altered by genetic engineering are called transgenic plants. E.g. Bt. Cotton, Bt-tomato, Flavr-saver variety of tomato.

Q.17 What are transgenic animals?

Ans.Animals that have had their DNA manipulated to possess and express an

extra (foreign) gene are known as transgenic animals. Transgenic rats,rabbits, pigs, sheep, cows and fish.

Q.18 What are cry genes?

Ans.These are the genes found in bacteria Bacillus thuringienis which produce an

insecticidal crystal protein called as cry protein.

Q.19 Mention two objectives of setting up GEAC by our government?

Ans.1. To make decisions regarding the validity of GM research.

2. To ensure the safety of introducing GM organisms for public services.

Q.20 State the role of C-peptide in human insulin.

Ans.The C-peptide is a short 31-amino acid polypeptide that connects insulin's A-

chain to its B-chain in the proinsulin molecule. It is also called as connecting

peptide. It also play important role in the synthesis of insulin.

A217

RECAPITULATION

INTRODUCTION

BIOTECHNOLOGY-The term Biotechnology was coined in year 1919 by an agricultural engineer Karoly Ereky, hence he is called as

father of biotechnology.Principles of biotechnology According to modern biotechnology the main principles of biotechnology are:

1. Genetic Engineering

2. Bioprocess Engineering

3. Recombinant DNA Technology

4.DNA Cloning

BIOTECHNOLOGICAL APPLICATION IN AGRICULTURE:

Production of GMO (Genetically modified organisms)

Transgenics plants are produced

Bt cotton- Tne genetically modified crop is called bt cotton as it contain Bt toxin

genes. Bt toxin produced certain proteins that kill insect

Formation of pest resistant plants using Agrobacterium vectors

Golden rice is transgenic variety which contain good quantity of vitamin A

BIOTECHNOLOGICAL APPLICATION IN MEDICINE:

In case of insulin production genes coding for human insulin are inserted into

bacteria produce insulin which is used for diabetic patient.Role of biotechnology in molecular diagnosis:

ELISA is based on principle of antigen and antibody interaction.

Gene therapy is technique of genetic engineering to replace faulty gene by normal healthy functional gene

PCR is done to amplify the content of DNA

Ethical issues:Introduction of trans genes from one species into another species violates the integrity of the species there is another biological damage it can accidently create new infectious agents

2-MARKS QUESTIONS ANSWER

Q1. Name the genetically engineered human insulin.

Ans. Humulin is the genetically engineered human insulin

Q2. What do you mean by Golden Rice?

Ans. Golden rice is a variety of rice produced through genetic engineering

to Bio synthesizes the precursors of beta-carotene. Golden rice has 23 times

Provitamin - A. The colour of golden rice is golden due to synthesis of Provitamin - A in entire grains.

Q3. Write brief account on recombinant Insulin.

Ans. In 1983 ELI LILLY an American company prepared two DNA Sequences corresponding to A and B chains of insulin such DNA Sequences

were introduced in E-coli plasmid when these bacteria produced chains than these chains were extracted and combined by disulphide bonds to form human insulin called Humulin.

Q4. Write the full form of the terms PCR and ELISA.

Ans. PCR- Polymerase Chain reaction. It is used to amplify the gene of interest.

ELISA- Enzyme Linked Immuno Sorbent Assay. It is used for early diagnosis of the disease and results can be obtained at very low

concentration of Pathogen in the body.

Q5. What is Green Revolution? Write the basic elements.

Ans. Many times increase in yield of food crops due to improved crop varieties, use of better management practices and use of agrochemicals.Green revolution had succeeded in tripling the yield of crops due to

(i)Improved crop varieties.

(ii) Use of better management practices.

(ii) Use of agrochemicals i.e. fertilizers, insecticides & pesticides.

Q6. What is Ti plasmid? Name the organism where it is found?

Ans. The Ti plasmid, which stands for tumor inducing, is a piece of DNA that occurs in soil inhabiting plant pathogenic bacterium Agrobacterium tumefactions which induces formation of cancerous Growth called crown gall tumour in several dicot plants.

Q7. What is a Gene Therapy? What is the cause of Adenosine De Aminase deficiency.

Ans. It is a technique of genetic engineering to replace a faulty gene by a normal healthy functional gene in humans. The cause of ADA deficiency is the defective autosomal gene. Thus, this particular gene does not synthesis

the enzyme adenosine deaminase.

Q8.What are transgenic plants? Give two examples of these plants.

Ans. These plants whose genes have been altered by genetic engineering are called transgenic plants.e.g. Bt.cotton,Bt-tomato

Flavr- Saur variety of tomato.

Nemotoda resistant tabacoo plants.

Q9. What are transgenic animajs? Give two examples of these animals.

Ans. Animals whose genes have been altered by genetic engineering are called transgenic animals e.g. rosie cow, transgenic mice, etc.

Q10. What is bio-technology? Name two important products of bio-technology.

Ans. Branch of microbiology concerned with the use of microbes, plants,animal cells and their components to generate useful products & services in industrial processes is called biotechnology important products.

(i) Biopharmaceuticals, anti-biotics, vaccines

(ii) Fermented beverage

Q11. What are Cry Proteins? Name an organism that produces it.

Ans. Cry proteins are encoded by genes named cry. They are produced in Bacillus Thuringienis. Cry proteins are toxic to insects and act as Insecticides.

Q12. Write note on Biopatent.

Ans. The patent being given to the Biological agents and their products are called biopatent. Biopatents are awarded for the following;

(i)Genetically modified microbes

(ii) Gene Sequence

(iii) GM plants & animals

(iv) Biotechnological procedures

Q13. How did the first transgenic cows Rosie differ from other cows with

respect to quality of milk?

Ans. Rosie produced human protein alpha-lactaalbumin enriched milk which is nutritionally a more balanced product for human babies.

Q14. What do you mean by Biopiracy?

Ans. Biopiracy refers to the use of bioresources lay multinational companies & other organizations without proper authorization from countries & people concerned.

Q15. Name two pest resistant plants produced bu using DNA recombinant Technology?

Ans. Bt Cotton, Bt Corn, Bt Brinjal are pest -resistant plants produced by DNA Recombinant technology.

Q16. Name any disease against which vaccine is developed by DNA Recombinant technology ?

Ans. Hepatitis B the disease against which vaccines are developed by DNA recombinant technology.

Q17. What is the full form of ADA in ADA deficiency?

Ans. Adenosine De Aminase Q18. Can you suggest a method to remove oil from seeds based on DNA Technology and chemistry of oil?

Ans. The oil in seed is composed of glycerol or fatty cids. Production of these

molecules is governed by particular gene. Such genes can be removed by using DNA technology. This will help in production of oil less seeds.

Q19. What are antigens?

Ans. Antigens are the substance that stimulates the immune system to

produce antibodies. Antigen can be bacteria, virus, or fungi that can cause infection and disease.

Q.20 Write note on GEAC.

Ans. GEAC stands for Genetic engineering approval committee. The Indian

Government has set up it for making decisions regarding the validity of modifications and safety of introducing genetically modified organisms for public services.

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RECAPITULATION

The applications of biotechnology include therapeutics, diagnostics, and

genetically modified crops for agriculture, processed food, bioremediation,waste treatment and energy production.

BIOTECHNOLOGICAL APPLICATIONS IN AGRICULTURE

Production of GMO (Genetically modified organisms).

Formation of pest resistant plants using Agrobacterium vectors

Transgenic plants are produced Bt cotton- The genetically modified crop is called Bt cotton as it contains Bt toxin genes. Bt toxin genes isolated from Bacillus thuringiensis which produces certain proteins that kill insects such as lepidopterans, coleopterans, and dipterans.

Golden rice is a transgenic variety of which contains good quantities of beta carotene.

BIOTECHNOLOGICAL APPLICATIONS IN MEDICINE

Formation of genetically modified insulin Known as humulin. In the treatment of a large number of Diabetes patients , humulin helps a lot.

ROLE OF BIOTECHNOLOGY IN MOLECULAR DIAGNOSIS

Gene therapy is the technique of genetic engineering to replace a faulty gene by a normal healthy functional gene.

ELISA is based on the principle of antigen-antibody interaction.Infection by pathogens can be detected by the presence of antigens or by detecting the antibodies synthesized against the pathogen.

PCR is done to amplify the content of DNA.

ETHICAL ISSUES:The Indian government has set up organisations such as GEAC (Genetic Engineering Approval Committee) which are authorised to make decisions regarding the validity of genetic modifications and safety of introducing genetically modified organisms for public services.

Biopiracy is use of bio resources by multinational companies and other

organization without proper authorization from the countries and people concerned without compensatory payment.

SHORT ANSWER TYPE QUESTIONS

Q 1.What is transgenic organism?

Ans: An organism which carries a foreign functional gene in its genome is termed

as transgenic organism.

Q2. Expand the following terms and mention one application of each a. PCR b. ELISA

Ans: a) PCR- Polymerase Chain Reaction. It is used to amplify the gene of interest.

b) ELISA- Enzyme Linked Immuno-Sorbent Assay. It is used for early diagnosis of disease and results can be obtained at very low concentration of pathogen in the body.

Q3: What are vaccines?

Ans- Vaccines are either dead or attenuated agents of disease which when injected into healthy person provide immunity to that particular disease.

Q4. What do you mean by Golden Rice?

Ans: Golden rice is a variety of rice produced through genetic engineering to

biosynthesize the precursors of beta- carotene. Golden rice has 23 times more

provitamin A. The colour of golden rice is golden yellow due to synthesis of

provitamin A in entire grains.

Q5. Write a note on Bt cotton.

Ans- It is genetically engineered plant. Bacillus thuringiensis is a naturally

occurring soil bacterium that produces a crystal protein that is toxic to insect

larvae. Crystal protein genes have been transferred into cotton plant, enabling the

cotton plant to produce its own pesticides against insects such as bollworm.

Q6: What are transgenic bacteria?

Ans- The bacteria whose genes have been treated by genetic engineering is called

transgenic bacteria. E.g. E.coli bacteria having human insulin gene.

Q7. Define antibiotic.

Ans- An antibiotic is an organic compound produced by a microorganism that

inhibits the growth of or kills another microorganism.

Q8: What are Cry proteins? Name an organism that produces it.

Ans- Cry proteins are encoded by genes named cry They are produced in

Bacillus thrugiensis. Cry proteins are toxic to insects and act as insecticides.

Q9: What is Ti-plasmid ? Name the organism where it is found?

Ans: The Ti-plasmid , which stands for tumour inducing , is a piece of DNA that

occurs in soil inhabiting plant pathogenic bacterium Agrobacterium tumefaciens

which induces formation of cancerous growth called in several dicot plants.

Q10: What are transgenic plants? Give two examples of these plants.

Ans- The plants whose genes have been altered by genetic engineering are called

transgenic plants.e.g. Bt cotton, Nematode resistant tobacco plants.

Q11: Name the genetically engineered human insulin.

Ans- Humulin is the genetically engineered human insulin.

Q12: Why did bacterial toxins not kill the bacteria but only the insects?

Ans- Bacillus thuringiensis is not killed by the bacterial toxins because toxic protein

exists as an inactive protoxin but if it is consumed by an insect it gets transformed into an active form of toxin due to alkaline pH of the alimentary canal of insect.

Q13: Name the technique based on the principle of antigen-antibody interaction used in the detection of a virus (HIV).

Ans- The technique based on the principle of antigen-antibody interaction used in the detection of a virus is ELISA (Enzyme linked Immuno-sorbent assay).

Q14: write short on GEAC.

Ans: GEAC stands for Genetic engineering approval committee. The Indian government has set up it for making decisions regarding the validity of genetic modifications and the safety of introducing genetically modified organisms for public services.

Q15: Write any two advantages of production of genetically modified crops.

Ans: Advantages of genetically modified crops:

1.Genetically modified crops have improved agronomic and other features such as

resistance to biotic and abiotic stresses.

2. Over ripening losses can be reduced. e.g.flavr savr tomato.

Q16: Write two applications of biotechnology?

Ans- 1. Treatment of diseases

2. Preparation of processed fortified food.

Q17: What do you mean by term Biopatent?

Ans- A patent is a right granted by a government to an inventor to prevent others

to make commercial use of such an invention. Patents granted for biological entities and the various products obtained from these organisms termed Biopatent.

Q18: What are transgenic animals? Give two examples of these.

Ans- Animals whose gene has been altered by genetic engineering are called transgenic animals.e.g. Rosie cow, transgenic mice.

Q19: What is biopiracy?

Ans- Biopiracy is use of bio resources by commercial and multinational companies and other organization without the appropriate authorization and permission from the countries and people concerned.

Q20. Distinguish between Cry and cry.

Ans- Cry stands for crystal of toxic protein whereas cry is the gene responsible for synthesis of toxin.

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INTRODUCTION

Students, in previous sheet you have learnt about various 2 marks questions but in

this revision sheet you will discuss important topics in the form of 3 marks framed question from chapter Biotechnology and its application. These questions will

help you in preparing for your Borad exams.

3- MARKS QUESTIONS:

Q.1 What do you mean by Genetically Modified Crops? Write any 4

advantagesof Genetically Modified Crops.

Ans: Genetically Modified Organisms (GMOs) are plants, animals, bacteria and

fungi whose genes have been altered by manipulation.Advantages of GMOs are as follows:

(1) Higher yield- Crop plants can be made to grow fast and produce very high yield

through genetic modification, example, Super wheat.

(2) Nutritional value- Vitamin A rich Rice and protein rich potato have been

developed through genetic engineering.

(3) Stresses tolerant- Genetically engineered plants can tolerate drought, frost,salinity, heavy metal toxicity.

(4) Disease resistance- Resistance to viral disease has been introduced in tobacco,

potato, tomato and rice. Cry gene from Bacillus thuringiensis has been introduced in

Bt cotton and Bt corn. Nematode infection has been overcome in some plants through RNA interference.

Q.2 List the disadvantages of production of genetically modified crops.

Ans: (1) Genetic pollution- Transgenes can be transferred from one plant to another plant, microbes and even animals. It shall disturb the genetic setup of organism and

cause ecological imbalance.

(2) Superweeds- Weedicide’s genes are being introduced into crop plants. Any of these crop plants can itself become a super weed.

(3) Super insecticide- Cry gene being introduced in crop plants can pass into wild

vegetation. Insects feeding on pollen and other parts will be killed resulting in the

destruction of pollinators and disseminators.

(4) Damage to environment- Harvesting leaves a lot of plant residue in the farm land

that would damage the biotic environment.

(5) Allergies: The products of transgenes are foreign to human body and animals feeding on transgenic. This may cause allergy and toxicity.

(6) Antibiotic resistance: Antibiotic resistance gene present in transgenic food

canbe picked up by bacteria present in human gut and transfer the same to pathogens.

Q.3 With an example, explain how biotechnology has been applied in each

ofthe following:

(i) in curing diabetes mellitus

(ii) in raising pest resistant plants

(iii) in producing more nutritionally balanced milk

Ans:(i) Diabetes mellitus- By producinginsulin or Humulin§ from transgenic

Escherichia coli.

(ii) Pest resistant plants- By m RNA silencing through the technique of RNA

interference which produces both sense and antisense RNAs

(iii) Transgenic cattle- Cattle have genes for milk proteins, growth hormones and

casein given high yield of nutritionally balanced milk with high protein content.

Q.4 What is RNA silencing? How is this strategy used to create pest

resistanceplants?

Ans: RNA silencing is a technique which involves silencing or disabling of specific mRNA due to complimentary ds RNA molecule that binds to and prevent translation

of MRNA.This strategy is used to prevent infection of roots of tobacco plants by nematode

Meloidegyne incognita. In this strategy, complementary ds RNA are produced

against specific mRNA. The source of this complementary RNA could be from an infection by viruses having RNA genome. Using Agrobacterium vector nematode

specific genes were introduced into host plants. The introduction of DNAwas such that it produced both sense and anti-sense RNA in the host cell. These two RNAs

being complementary to each other formed a double stranded RNA that initiated

RNAi and thus silenced specific mRNA of the nematode. The consequencewas that parasite could not survive in transgenic host.

Q.5 What are cry proteins? Name an organism that produces them. How

manhas exploited proteins to his benefit?

Ans: Cry proteins are a group of toxic proteins which are highly poisonous to

different types of insects, example, tobacco bud worm, army worm, beetles, flies and

mosquitoes.The organism producing Cry proteins is Bacillus thuringiensis. The genes controlling their formation were called cry genes. The bacterium produces the protein as

endotoxin in protoxin crystalline state.

Two cry genes have been incorporated in cotton (Bt cotton) while one has been

introduced in corn. As a result, Bt cotton has become resistant to boll worms while

Bt corn has developed resistance to corn borer.

Q.6 (i) Name the pest that destroys the cotton bolls.

(ii) Explain the role of Bacillus thuringiensis in protecting the cotton crop

against the pest to increase the yield.

Ans: (i) The pest that destroys the cotton balls are cotton boll worms and cotton

borer.

(ii) Bt cotton is created by using some strains of a bacterium, Bacillus

thuringiensis. This bacterium produces protein, that kill certain insects such as

lepidopterans (tobacco budworm and armyworm), coleopterans (beetles) and

dipterans (flies and mosquitoes). Bacillus thuringiensis forms protein crystals

during a particular phase of their growth. These crystals contain a toxic insecticidal

protein. Bt toxin protein exist as inactive protoxin, but once an insect ingests the

inactive toxin, it is converted into an active form due to the alkaline pH of the gut

which solubilize the crystals. The activated toxin binds to the surface of midgut

epithelial cells and create pores that cause cell swelling and lysis leading to death

of insect. Specific Bt toxin genes were isolated from Bacillus thuringiensis and

incorporated into several crop plants. Most Bt toxins are insect-group specific.

Hence, the toxin is coded by a gene named cry. For example, the proteins encoded by the genes cry | Ac and cry | Ab control the cotton boll worms and cry |Ab controls corn borer.

Q.7 (i) Explain the effect of deletion of the gene for ADA in an individual.

(ii) How does the gene therapy help in this case?

Ans. (i) Deletion of the gene for ADA in an individual leads to ADA deficiency

disorder. Adenosine Deaminase (ADA) enzyme is crucial for immune system to function.

(ii) Gene therapy is helpful in case of ADA deficiency.Hereditary disease can be corrected by gene therapy. It is a collection of methods that allows correction or replacement of defective gene.The first gene therapy was given in 1990 to a 4 years old girl with Adenosine Deaminase (ADA) deficiency. It is caused due to the deletion of gene for adenosine deaminase.The treatment involves following steps:

(a) Lymphocytes from the blood of patient are grown on culture outside the body. A functional ADA, c DNA (using a Retro viral vector) is then introduced into teelymphocytes.

(b) Such genetically engineered lymphocytes are returned to the blood of patient.

Q.8 What is molecular diagnosis? Explain.

Ans: Molecular diagnosis: Effective treatment of diseases require early diagnosis

and understanding of their pathophysiology.

Early diagnosis of diseases is not possible by conventional techniques like serum

and urine analysis, sputum and stool analysis etc.Modem techniques like rDNA, PCR, ELISA and RIA are used for early diagnosis.

Infection of pathogens (viruses, bacteria etc.) is suspected only when they produce disease symptoms.

By this time the pathogens have multiplied in the body.

But by PCR, amplification of nucleotide is possible and thus it is possible to

detect bacteria, virus even ats very low concentrations. e.g.:HIV detection, genetic disorders.Modem techniques can also help in the identification of mutated genes in humans

A single stranded DNA or RNA, tagged with a radioactive probe is allowed to hybridize with its complementary DNA in a clone of cells, followed by detection by using autoradiography.

The clone having mutated gene will not appear in the photographic film because the probe will not have complementary nitrogen base with the mutated gene.

In ELISA test the infection of pathogen (viruses, bacteria, fungi, mycoplasma

like organism) can detect the antibodies synthesized against the pathogen.

Q.9 (i) Name the host plant and its part that Meloidogyne incognita infects.

(ii) Explain the role of Agrobacterium in the production of dsRNA in the host plant.

Ans. (i) The nematode Meloidogyne incognita infects the roots of tobacco plants.

(ii) The Agrobacterium are used as vectors carrying nematode specific genes

to be introduced in host plant. These genes when expressed inside host plant

produces sense and anti-sense RNA strands, complementary to nematode’s functional MRNA. This binding result in formation of double stranded RNA and inhibiting or silencing the translation of RNA specified. This process is called RNA interference.

Q.10 In the given figure, Form (A) and Form (B) represent different forms of aproteinaceous hormone secreted by pancreas in mammals.

(a) What type of bonding is present between chains of this hormone?

(b) What are these form (A) and form (B). How these forms differ from each

other?

Ans. (a) Disulphide bonds

(b) Form (A) — Proinsulin

Form (B) — Mature insulin. Proinsulin contains an extra stretch called C- peptide

which is absent in mature insulin.

B peptide chains of human insulin and introduced them in plasmid E. coli

to produce insulin chains. Chains A and B were produced separately,extracted and combinedby creating disulphide bonds to form insulin.

Q.11 What are the steps involved in synthesis of genetically engineeredinsulin.

Ans. Steps involved in Insulin production are:-

1. For the synthesis of Insulin, RNA is extracted from B- cells of islets of

Langerhans of pancreas.

2. With the help of enzyme Reverse transcriptase, single stranded DNA

complementary to mRNA is synthesized. Second strand of DNA complementary to first is synthesized with enzyme DNA polymerase.

3. The two strands of copy DNA is joined to plasmid by using an enzyme called

terminal transferase.

4. The two ends of DNA get annealed by enzyme called ligase thus ends of inserted DNA & plasmid are sealed & a new circular plasmid is formed. This is a molecule of recombinant DNA.

5. This recombinant DNA is then inoculated in a new bacterial cell of E-coli & inserted in a bacterial gene after having cut by restriction enzyme.

6. After proper expression of genes the bacterial cells of both cultures are lysed

with appropriate chemicals. The fragments of insulin are then separated from enzyme by Cyanogen Bromide.

Q.12 Some multinational companies and other organizations are using bioresources for commercial benefits, without proper authentication andcompensation to concerned authorities.

(a) Give the term for this unauthorized act.

(b) Suggest any two ways to get rid of this.

Ans. (a) Biopiracy

(b) (i) Benefits of bioresources should be shared between developed and developing nations.

(ii) Laws should be developed to prevent unauthorsied exploitation of them bioresources.

Q.13 Describe with example, why transgenic animals are produced?

Ans. Transgenic animals are produced for following purposes: -

1. To allow the study of how genes are regulated & how they affect normal function of body & its development e.g. Information obtained about biological role of insulin like growth factor.

2. To increase our understanding on how genes contribute to development of diseases.

3. To produce useful biological compounds by introducing a portion of DNA that codes for that product from other organisms, e.g. aa-1 antitrypsin, a protein used to treat emphysema.

4. For testing the safety of vaccine eg. Polio vaccine in transgenic mice.

5. To test the toxicity of drugs.

Q.14 (i) Some crop plants are modified genetically by manipulating their

genes.How are they made beneficial?

(ii) What is Golden rice? What is its advantage?

Ans: (i) More tolerant to abiotic stresses; pest resistant; reduction in post harvest

losses; increased nutritional value of food.

(ii) Golden rice is a transgenic variety of rice which contains a gene which codes for Vitamin A precursor. This variety has green yellow colored grains and is rich in Vitamin A & thus nutritionally very advantageous.

Q.15(i) Human insulin when synthesized in the body needs to be processed before it can act. Explain giving reasons.

(ii) How is the insulin produced from recombinant DNA technology different from that produced by the functional human insulin gene?

Ans: (i) Human insulin when initially synthesised in human body consists of three

peptide chains- A, B and C. The C-peptide is an extra stretch of amino acids joining

the A and B-chains. This is called Proinsulin or Prohormone.It undergoes processingor splicing to release the functional mature insulin that can carry out its normal functions. During processing, the C-peptide is removed. Only A and B-chainscontribute to form the functional insulin.

Q.16 (i) Why is Bacillus thuringiensis considered suitable for developing GM plants?

(ii) Explain how it has been used to develop GM crops.

Ans. (ii) Bacillus thuringiensis produces a protein, which is toxic to the larvae of

insects like boll worm, budworm, flies, beetle, etc. Bt toxin gene is cloned from the

bacteria and has been expressed in plants to provide resistance against insects

without the need for chemical insecticides.

(ii) The method for developing GM plants involve:

cry gene coding for proteins has been isolated using restriction enzymes. They are

cloned in the vectors and then introduced into desired crop plants. The different

types of cry genes that code for insect-specific cry proteins are cry l|Ac and cry IlAb that control cotton bollworm. crylAb controls corn borer.

(iii) The transgenic plants, i.e., Bt cotton, Bt corn, Bt rice, produce the protein in their cells and express insect pest resistance.

Q.17 What are transgenic bacteria? Illustrate using any one example.

Ans: Transgenic bacteria are microbes carrying clones of foreign genes.Transgenic bacteria are being employed for a variety of functions.

(1) Biochemical factories- Microbes are used for producing important biochemicals. It is an old use of microbes. Traditionally microbes have been synthesizing alcohol, enzymes, steroids, organic acids and antibiotics. Transgenic bacteria have added anumber of biochemicals to this list, example, human insulin,growth hormone, tissue plasminogen factor, erythropoietin, fertility hormone, lung surfactant protein, blood clotting factors, interferons, interleukin etc. Some genetically modified bacteria have been made to augment traditional functions,

example, Pseudomonas putida for alcoholic fermentation.

(2) Environmental protection- Microbes have been genetically changed to help in

cleaning the polluted environment, example, Pseudomonas putida for cleaning oil

spills, Pseudomonas for removing heavy metal pollutants, Acetobacter aerogens for

decomposition of DDT and Flavobacterium for decomposition of 2,4-D.

Q.18 Mention the advantages & disadvantage of production of Genetically modified organisms?

Ans. ADVANTAGES OF PRODUCING GMOs:

1. GM crops produce desired phenotypic traits in crop plants. The genes responsible for production of specific proteins are inserted into GMcrops.These crops then produce that specific protein.

2. Transgenic crops synthesizes new end product of specific biochemical pathway.

3. These crops also help in preventing expression of existing native Gene.

DISADVANTAGES OF PRODUCING GMOS:

1. Transgenic crops may endanger wild & native species.

2. GM crops may cause health problems by supplying allergens.

3. GM crops may damage to the natural environment.

Q.19 (i) List the three molecular diagnostic techniques that help to detect pathogens from suspected patients.

(ii) Expand ELISA. On what principle is ELISA test based? List two ways by which an infection can be detected by this test.

Ans: (i) Molecular diagnostic techniques to pathogens are as follow:

(a) Polymerase Chain Reaction (PCR).

(b) Recombinant DNA technology.

(ii) ELISA — Enzyme Linked Immuno Sorbent Assay. ELISA is based on antigen-antibody interaction.The two ways to detect the presence of infection or disease by ELISA are as follow:

(a) The presence of antigens (proteins, glycoproteins, etc) is detected.

(b) Antibodies produced against the pathogen are detected.

Q.20 Diagrammatically represent the experimental steps in cloning and

expressing a human gene (say the gene for growth hormone) into a bacterium like E. coli?

Ans: A technique of producing multiple identical copies of a particular template DNA

is known as DNA cloning which involves the usage of a vector that carries the

particular foreign DNA fragment into the host cell.The working of cloning and transfer of gene for growth hormone into E.coli can bedepicted as given below:

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RECAPITULATION

Biotechnology is a broad area of biology, involving the use of living systems and

organisms to develop or make products. Biotechnology has given to humans

several useful products by using microbes, plant, animals and their metabolic

machinery.

APPLICATION OF BIOTECHNOLOGY

The applications of biotechnology include therapeutics, diagnostics, and

genetically modified crops for agriculture, processed food, bioremediation, waste

treatment, and energy production.Recombinant DNA technology has made it possible to engineer microbes, plants and animals such that they have novel capabilities. Genetically Modified Organisms have been created by using methods other than natural methods to transfer one or more genes from one organism to another, generally using

techniques such as recombinant DNA technology.

BIOTECHNOLOGICAL APPLICATION IN AGRICULTURE

The plants in which foreign genes have been introduced trough genetically engineering are called transgenic plants.Advantages of transgenic plants GM plants have been useful in

increasing crop yields

pest resistant crops

reduce post-harvest losses and make crops more tolerant of stresses.Disadvantages of transgenic plants

Environmental hazards

Human health risks like allergies

BIOTECHNOLOGICAL APPLICATIONS IN MEDICINE

Recombinant DNA technological processes have made immense impact in the area of healthcare by enabling mass production of safe and more effective therapeutics. Recombinant pharmaceuticals or therapeutics is created by inserting genes from one species into a host species, often yeast or bacteria. For example,In case of insulin production, genes coding for human insulin are inserted into

bacteria. Bacteria produce insulin, which is harvested and used for diabetic patients. Human insulin is made in bacteria yet its structure is absolutely identical to that of the natural molecule.

ADVANTAGES OF RECOMBINANT THERAPEUTICS:

*Recombinant therapeutics never induce unwanted immunological responses as is

common in case of similar products isolated from non-human resources or other

animals which contain other proteins and generate immune responses.

Recombinant therapeutics is produced in very large amounts and also very easy to produce. *These drugs are very effective with no any kind of side effects.

Production of these drugs are time and money saving.30 recombinant therapeutics have been approved for human use on word level but in India 12 of these are approved and presently being marketed.

TRANSGENIC ANIMALS

Transgenic animals are also used to understand how genes contribute to the

development of a disease by serving as models for human diseases, such as cancer, cystic fibrosis, rheumatoid arthritis and Alzheimer's.

GENE THERAPY

GENE THERAPY is a collection of methods that allows correction of a gene defect

that has been diagnosed in a child/embryo. Genes are inserted into a person's

cells to treat a disease. Correction of a genetic defect involves delivery of a normal

gene into the individual or embryo to take over the function and compensate for the

non-functional gene. The first clinical gene therapy was given in 1990 to a 4-year

old girl with Adenosine De Aminase (ADA) deficiency.

ETHICAL ISSUES IN BIOTECHNOLOGY

I. Introduction of a transgene from one species into another species violates

the integrity of species.

Il. | Biotechnology may lead to unforeseen risks to the environment, including

risk to biodiversity. it can disturb the existing ecological balance.

Ill. Utilization of animals In biotechnology causes great suffering to them.

IV. There is another biological damage .it can accidentally create new infectious agents.

V. | When animals are used for production of pharmaceutical proteins they are virtually reduced to the status of a factory.

Revision test sheet Time allowed: 2hrs. Max. Marks: 35

Important Instructions:

1. There are a total of 17 questions and four sections in the question paper.All questions are compulsory.

2. Section A contains question numbers 1 to 5 very short answer type questions of one mark each.

3. Section B contains question numbers 6 to 13, short answer type questions of two marks each.

4. Section C contains question numbers 14 to 16, short answer type II questions of three marks each.

5. Section D contains question number 17, long answer type question of five

marks. There will be internal choice in this question.

6. There is no overall choice in the question paper

Section-A

Each question carries 1 mark

Q1. What are DNA vaccines?

Q2. Name the bacterium that is first used as bio pesticide?

Q3. What are the transgenic animals?

Q4. Name the genetically engineered insulin.

Q5. What is the full form of ADA in ADA deficiency?

Section-B

Each question carries 2 marks

Q6. Name two pest-resistant plants produced by using recombinant DNA technology.

Q7. Which vaccine was being tested on mice?

Q8. Name any disease against which vaccine is developed by Recombinant DNA technology.

Q9. What is the difference between ‘cry’ & ‘CRY’?

Q10. What are the two methods for correcting ADA deficiency in a child?

Q11. What is Golden rice? What is its advantage?

Q12. The first transgenic cow produced human protein-enriched milk. Name the cow and the protein found in milk.

Q13. What do you mean by “Biopiracy” Give an example?

Section-C

Each question carries 3 marks

Q14. What do you understand by Molecular farming?

Q15. Describe some transgenic plants and their useful characters.

Q16. What are the advantages and disadvantages of transgenic plants?

Section-D

Each question carries 5 marks

17. Describe the biotechnical applications in agriculture.

18. How biotechnology is helpful in the field of medicine.

ANSWERS

Section-A

A1. Genetically engineered vaccines are called DNA vaccines.

A2. Bacillus thuringiensis.

A3. These are the animals, whose genome has been altered by introduction of an extra (foreign) gene by manipulation.

A4. Humulin.

Ad. Adenosine De Aminase (ADA) deficiency.

Section-B

A6. Bt Cotton, Bt Corn, BtBrinjal are pest-resistant plants produced by using

recombinant DNA technology.AT. Polio vaccine was being tested on mice.

A8. Hepatitis B is the disease against which vaccines are developed by Recombinant DNA technology.

AQ. cry is the gene that codes for Bt-toxin which is an insecticidal protein while CRY is the protein coded by cry genes.

A10. Enzyme replacement therapy and bone marrow transplantation with a functional ADA enzyme.

A11. Golden rice is a genetically modified type of rice rich in Vitamin A. This

type has green-yellow grains and is high in Vitamin A, making it nutritionally

beneficial.

A12. Rosie, is the first transgenic cow, it produced human protein-enriched milk. Alpha-lactalbumin is the protein found in milk.

A13. Biopiracy is defined as the use of bio-resources by multinational corporations and other organizations without proper authorization from the countries and people concerned. For example, Basmati rice grown in India is known for its distinct flavor and aroma, but Basmati rice was patented by an

American company under a US patent.

Section-C

A14. Molecular farming :The method of manufacturing drugs in transgenic

animals is called molecular farming or molecular pharming. The transgenic

animals can revolutionize the pharmaceutical industry by providing large quantities of valuable drugs.

A16. The plants in which foreign genes have been introduced trough genetically

engineering are called transgenic plants.

Advantages of transgenic plants:

GM plants have been useful in

increasing crop yields

pest resistant crops

reduce post-harvest losses and make crops more tolerant of stresses.

Disadvantages of transgenic plants:

Environmental hazards

Human health risks like allergies

Section-D

A17. Agriculture is a major sector for the production of food materials and

development of the nation also. But, now a day we are facing a huge problem with

food production for the need of growing human population. There are three possible options for enhancement of crop production.

1. Agro-chemical based agriculture, by using chemical fertilizers and pesticides.

2. Organic agriculture, by using organic or bio fertilizers and bio pesticides.

3. Genetically engineered crop based agriculture, in which the crops are

genetically modified to enhance the food production and to make them insect, pest

resistant.The first two options bring green revolution, but yet it was not enough to feed the growing human population. Then scientists have decided that use of genetically

modified crops is a possible solution.

The latest Recombinant DNA technology or Genetic Engineering plays an important role in agriculture. This technology involved transfer of one or more genes (i.e. DNA fragment) from one plant to another. The plant in which foreign genes has been introduced is called transgenic plant. Now these transgenic plants are used in agriculture for the following reasons:

1. Improvement In Photosynthesis - RUBISCO Is an important enzyme of photosynthesis which can act both on carbon dioxide and oxygen. When this enzyme combine with CO2 it performs C3 cycle in which carbon dioxide is

fixed into glucose. The property of any enzyme Combining with COz is called

carboxylase and combining with oxygen is called oxygenase. Whenever RUBISCO combines with oxygen, it performs C2 cycle also known as photorespiration. C2 cycle is a wasteful cycle for plants because in this cycle

plants starts losing their fixed carbon. The RUBISCO can act as carboxylase as well as oxygenase also. When it is acting as carboxylase, it is performing C3 cycle and when it is acting as oxygenase, it is performing C2 cycle. It is now possible to isolate the RUBISCO gene, modify it accordingly and reinsert into plants to minimise the loss due to photorespiration. The scientists removes the oxygenase property of RUBISCO with the help of genetic engineering and retains its carboxylase property. With this, scientists

are successful to minimise the loss of fixed carbon.

2. Insect - Pest Resistant In Transgenic Plants - The day is not far off wnen

our crop plants will resist all kind of pests. Several such transgenic plants

have been developed. Bacillus thuringiensis is a bacteria which produces a toxin protein which is a crystal protein which can kill a number of insects.This protein is also known as cry protein. Bt, toxin genes were isolated from

B. thuringiensis and added to several crop plants. Such plants can resist many kind of pests. One such plant is Bt. cotton in which Bt. Toxin has been used as a biological insecticide. This type of cotton is resistant to the boliworm. Tomatoes with Bt. Gene are completely protected from an attack of caterpillars. Similarly GM tobacco with Bt gene is protected from hornworms.

3. Herbicide tolerance - Herbicides are the chemicals which kill the weeds.When these Chemicals are sprayed in the field, they not only kill the weeds but also damage the crop plants. In recent years, genetic engineering has helped to develop such transgenic crop plants which are resistant to herbicides, so that they are not damaged when farmers Spray herbicides in the fields. Some herbicide resistant crops produced by genetic engineering are - tomato, potato, tobacco, soyabean, cotton, corn, oilseed, sugar beet,

etc.

A118. Viodern biotechnology is making a great contribution to medicine, both in the diagnosis of diseases and in the development of Pharmaceutical products for disease therapy. One purpose of DNA technology is to identify the gene that's mutation, causes genetic diseases. Discovery can help to

find ways for diagnosing, treating and preventing mutation. So this technology is very helpful in diagnosing, treating and preventing the mutation for helping the future generations to avoid the particular genetic disorders.

1. Polymerase chain reaction(PCR):PCR is a technique by which any piece of DNA can be quickly amplified(copied many times) Without using cells. The DNA is incubated in a

test tube with a special kind of a DNA polymerase enzyme, a mixture of deoxyribonucleotide for use as raw material, and short piece of synthetic single stranded DNA to serve as primers for DNA synthesis. PCR can make billions of copies of a DNA segment in a few hours. DNA technology can help identify individuals with genetic disorders before the appearance of

symptoms, even before birth.

2. Pharmaceutical products:Many Pharmaceutical products have been produced by using DNA technology. these products are mostly proteins. The gene for a desired

protein is transferred into a bacterium, yeast or other kind of cell which is

easy to grow in a culture. Such host cells produce large quantities of protein

in a short time.

a. Hormones - Human insulin and human growth hormone are earliest example.

(i) Insulin — Insulin is a hormone, produced by human body. Insulin converts glucose into glycogen which is stored in our liver. In the deficiency of insulin,glucose will not be converted into glycogen and glucose level of blood will increase and it will develop the disease diabetes. When human body is not

producing insulin then external insulin was given. Insulin is an important lifesaving drug for diabetic patients. Earlier, the insulin was obtained from the pancreas of cattle and pigs, and this is not sufficient as per the need of growing number of diabetic patients. Injections of this insulin have some side effects also. Scientists were looking for some alternative source of human

insulin. The search for new source was soon fulfilled by Recombinant DNA technology. The insulin was prepared with the help of bacteria using this technology.

Production of genetically engineered insulin:

Insulin is made up of two short polypeptide chains; A and B which are linked together by disulphide bridges. Insulin is synthesised as Pro hormone. It needs processing to become mature and functional.

It contains an extra stretch called C — peptide. This is removed during

maturation of insulin.

Genetic engineers took out the human gene responsible for synthesis of insulin and inserted it into the DNA chain of suitable vector, E. coli bacterial cell.

Chain A and B were produced separately, extracted and combined by creating disulphide bonds to form human insulin called Humulin. It was the first genetically engineered human insulin.

(ii) Human_growth hormone - Human Growth Hormones (HGH) has proved highly useful to numerous children born with HYPOPITUITARISM, a form of dwarfism caused by under secretion of the HGH by the anterior pituitary. The HGH

may also have other uses, such as healing of injuries.Milch cows are injected with Bovine Growth Hormone (BGH), formed in which

Genetically Engineered Microorganisms (E. coli), to raise milk production. BGH

also improve beef yield in cattle.

3. Gene Therapy — It is a collection of methods that allows correction of a gene

defect that has been diagnosed in a child/embryo. The disorders can be due to the

absence of a gene or due to non-functional gene which not able to work properly.

With the help of gene therapy, the correct gene is added into the body of patient or

embryo.