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CHANDRA SEKHAR ACADEMY

PURI-KONARK MARINE DRIVE ROAD, BALIGHAI, PURI, ODISHA

PURI-KONARK MARINE DRIVE ROAD, BALIGHAI, PURI, ODISHA 2018 – 2019 AN INVESTIGATORY PROJECT ON APPLICATIONS OF

2018 2019

AN INVESTIGATORY PROJECT ON

APPLICATIONS OF BIOTECHNOLOGY

AN INVESTIGATORY PROJECT ON APPLICATIONS OF BIOTECHNOLOGY SUBMITTED BY ASWESHA SARANGI CLASS – XII SUBJECT –

SUBMITTED BY

ASWESHA SARANGI CLASS XII SUBJECT BIOLOGY

SUBMITTED TO

PRIYAMBADA PATTANAIK

Contents

1. Introduction

2. History

3. Biotechnology in Agriculture

4. Genetically Modified Crops

RNA Interference (RNAi)

Bt toxin

5. Bt cotton

6. Biotechnology in Medicine

7. Genetically engineered insulin (Humulin)

8. Gene therapy

9. Conclusion

10.Bibliography

AKNOWLEDGEMENT

I am overwhelmed in all humbleness and gratefulness to acknowledge my depth to all those who have helped me to put these ideas, well above the level of simplicity and into something concrete.

I would like to express my special thanks of gratitude to my biology teacher,

PRIYAMBADA PATTANAIK as well as our Principal Miss. Amita Pattnaik who gave me the golden opportunity to do this wonderful project on the topic “Applications of Biotechnology”, which also helped me in doing a lot of research and I came to know about so many new things. I am really thankful to them.

Any attempt at any level can’t be satisfactorily completed without the support and guidance of my Parents and Friends who helped me a lot in gathering different information, collecting data and guiding me from time to time in making this project, despite of their busy schedules, they gave me different ideas in making this project unique. I am thankful to them too.

I am making this project not only for marks but to also increase my knowledge Thanking you

ASWESHA SARANGI CLASS - XII

I am making this project not only for marks but to also increase my knowledge Thanking

CERTIFICATE

This is to certify that ASWESHA SARANGI of class XII of CHANDRA SEKHAR ACADEMY has successfully completed the investigatory project on the topic “APPLICATIONS OF BIOTECHNOLOGY” under the guidance of PRIYAMBADA PATTANAIK during the session 2018-19 in the partial fulfilment of Biology Practical Examination conducted by CENTRAL BOARD OF SECONDARY EDUCATION (AISSCE).

Teacher’s Sign

Principal’s Sign

Student’s Sign

External’s Sign

INTRODUCTION

What is Biotechnology?

Biotechnology is the use of living systems and organisms to develop or make products, or "any technological application that uses biological systems, living organisms or derivatives thereof, to make or modify products or processes for specific use. At its simplest, biotechnology is technology based on biology biotechnology harnesses cellular and bio molecular processes to develop technologies and products that help improve our lives and the health of our planet. We have used the biological processes of microorganisms for more than 6,000 years to make useful food products, such as bread and cheese, and to preserve dairy products.

such as bread and cheese, and to preserve dairy products. Modern biotechnology provides breakthrough products and

Modern biotechnology provides breakthrough products and technologies to combat debilitating and rare diseases, reduce our environmental footprint, feed the hungry, useless and cleaner energy, and have safer, cleaner and more efficient industrial manufacturing processes.

Biotech is helping to heal the world by harnessing nature's own toolbox and using our own genetic makeup to heal and guide lines of research by:

Reducing rates of infectious disease

Saving millions of children's lives

Changing the odds of serious, life-threatening conditions affecting millions around the world

Tailoring treatments to individuals to minimize health risks and side effects

Creating more precise tools for disease detection

Combating serious illnesses and everyday threats confronting the developing world.

BIOTECHNOLOGY IN EARLY DAYS

Throughout the history of agriculture, farmers have inadvertently altered the genetics of their crops through introducing them to new environments and breeding them with other plants - one of the first forms of biotechnology.

These processes also were included in early fermentation of beer. In brewing, malted grains (containing enzymes) convert starch from grains into sugar and then adding specific yeasts to produce beer. In this process, carbohydrates in the grains were broken down into alcohols such as ethanol. Fermentation was also used in this time period to produce leavened bread. Although the process of fermentation was not fully understood until Louis Pasteur's work in 1857, it is still the first use of biotechnology to

convert a food source into another form.

of biotechnology to convert a food source into another form. For thousands of years, humans have

For thousands of years, humans have used selective breeding to improve production of crops and livestock to use them for food. In selective breeding, organisms with desirable characteristics are mated to produce offspring with the same characteristics. For example, this technique was used with corn to produce the largest and sweetest crops.

Biotechnology has also led to the development of antibiotics. In 1928, Alexander Fleming discovered the mould Penicillium.

BIOTECHNOLOGY IN AGRICULTURE

Genetically Modified Crops

BIOTECHNOLOGY IN AGRICULTURE Genetically Modified Crops Genetically modified crops o r “GM crops” or “biotech

Genetically modified crops or “GM crops” or “biotech crops” are plants used in agriculture, the DNA of which has been modified with genetic engineering techniques. In most cases the aim is to introduce a new trait to the plant which does not occur naturally in the species.

Examples in food crops include resistance to certain pests, diseases, stressful environmental conditions, resistance to chemical treatments, reduction of spoilage, or improving the nutrient profile of the crop. Examples in non-food crops include production of pharmaceutical agents, bio fuels, and other industrially useful goods, as well as for bioremediation.

Plants and crops with GM traits have been tested more than any other cropswith no credible evidence of harm to humans or animals. In fact, seeds with GM traits have been tested more than any other crops in the history of agriculture with no credible evidence of harm to humans or animals.

Genetic modifications have:

1. Made crops more tolerant to abiotic stresses (cold, drought, salt, heat).

2. Reduced reliance on chemical pesticides (pest resistant crops).

3. Helped to reduce post harvest losses & enhanced the nutritional value of the foods.

RNA Interference (RNAi)

RNA interference (RNAi) is a method of blocking gene function by inserting short sequences of ribonucleic acid (RNA) that match part of the target gene’s sequence,

thus no proteins are produced. RNAi has the potential to become a powerful therapeutic approach toward targeted and personalized medicine. RNAi has provided a way to control pests and diseases, introduce novel plant traits and increase crop yield. Using RNAi, scientists have developed novel crops such as nicotine-free tobacco, non-allergenic peanuts, decaffeinated coffee, and nutrient fortified maize among many others.

Mechanism of RNA interferences as understood is that it comes into play when a double stranded RNA is introduced either naturally or artificially in a cell. An endo ribonuclease enzyme cleaves the long dsRNA into small pieces of RNA. The small pieces could be mi RNA or si RNA depending upon the origin of long dsRNA i.e. endogenous or exogenous

upon the origin of long dsRNA i.e. endogenous or exogenous respectively. A double stranded RNA may

respectively.

A

double stranded RNA may be generated by either RNA dependent RNA polymerase or bidirectional transcription of transposable elements or physically introduced.

There

opportunities for

are several

the

applications of RNAi in crop science for its improvement

such as stress tolerance and enhanced nutritional level.This knockdown technology may be useful in inducing early flowering, delayed ripening, delayed senescence, breaking dormancy, stress- free plants, overcoming self-sterility, etc.

RNA interference (RNAi) has recently been demonstrated in plant parasitic nematodes. It is a potentially powerful investigative tool for the genome-wide identification of gene function that should help improve our understanding of plant parasitic nematodes. RNAi should help identify gene and, hence, protein targets for nematode control strategies. Prospects for novel resistance depend on the plant generating an effective form of double-stranded RNA in the absence of an endogenous target gene without detriment to itself. These RNA molecules must then become available to the nematode and be capable of ingestion via its feeding tube. If these requirements can be met, crop resistance could be achieved by a plant delivering a dsRNA that targets a nematode gene and induces a lethal or highly damaging RNAi effect on the parasite.

Bt Cotton

Bt cotton is a genetically modified organism (GMO) cotton variety, which produces an insecticide to bollworm. Strains of the bacterium Bacillus thuringiensis produce over 200 different Bt toxins, each harmful to different insects. Most notably, Bt toxins are insecticidal to the larvae of moths and butterflies, beetles, cotton bollworms and ghtu flies but are harmless to other forms of life. The gene coding for Bt toxin has been inserted into cotton as a transgene, causing it to produce this natural insecticide in its tissues. In many regions, the main pests in commercial cotton are lepidopteran larvae, which are killed by the Bt protein in thegenetically modified cotton they eat. This eliminates the need to use large amounts of broad-spectrum insecticides to kill lepidopteran pests. This spares natural insect predators in the farm ecology and further contributes to non insecticide

and further contributes t o non insecticide pest management. Bt cotton is ineffective against many cotton
and further contributes t o non insecticide pest management. Bt cotton is ineffective against many cotton

Bt cotton is ineffective against many cotton pests such as plant bugs, stink bugs, and aphids; depending on circumstances it may be desirable to use insecticides in prevention.

Mechanism:

Bt cotton was created through the addition of genes encoding toxin crystals in the Cry group of endotoxin. When insects attack and eat the cotton plant the Cry toxins are dissolved due to the high pH level of the insects stomach. The dissolved and activated Cry molecules bond to cadherin-like proteins on cells comprising the brush border molecules. The epithelium of the brush border membranes separates the body cavity from the gut whilst allowing access for nutrients. The Cry toxin molecules attach themselves to specific locations on the cadherin-like proteins present on the epithelial cells of the midge and ion channels are formed which allow the flow of potassium. Regulation of potassium concentration is essential and, if left unchecked, causes death of cells. Due to the formation of Cry ion channels sufficient regulation of potassium ions is lost and results in the death of epithelial cells. The death of such cells creates gaps in the brush border membrane.

Advantages:

Bt cotton has several advantages over non Bt cotton. The important advantages of Bt cotton are briefly :

Increases yield of cotton due to effective control of three types of bollworms, viz. American, Spotted and Pink bollworms.

Insects belonged to Lepidoptera (Bollworms) are sensitive to crystalline endotoxic protein produced by Bt gene which in turn protects cotton from bollworms.

Reduction in pesticide use in the cultivation of Bt cotton in which bollworms are major pests.

Reduction in the cost of cultivation and lower farming risks.

Reduction in environmental pollution by the use of insecticides rarely.

Bt cotton exhibit genetic resistance or inbuilt resistance which is a permanent type of resistance and not affected by environmental factors. Thus protects crop from bollworms.

Bt cotton is ecofriendly and does not have adverse effect on parasites, predators, beneficial insecticides and organisms present in soil.

It promotes multiplication of parasites and predators which help in controlling the bollworms by feeding on larvae and eggs of bollworm.

No health hazards due to rare use of insecticides.

Bt

early in

maturing as compared to non Bt cotton.

cotton

are

• No health hazards due to rare use of insecticides. • Bt early in maturing as

Disadvantages:

Bt cotton has some limitations

High cost of Bt cotton seeds as compared to non Bt cotton seeds.

Effectiveness up to 120 days, after that the toxin producing efficiency of the Bt gene drastically reduces.

Ineffective against sucking pests like jassids, aphids, whitefly etc.

Bt cotton in India:

Bt cotton is supplied in India's Maharashtra state by the agribiotechnology company, Mahyco, as the distributor.

The use of Bt cotton in India has grown exponentially since its introduction. Recently India has become the number one global exporter of cotton and the second largest cotton producer in the world. India has bred Bt-cotton varieties such as Bikaneri Nerma and hybrids such as NHH-44, setting up India to benefit now and well into the future.

India’s success has been subject to scrutiny. Monsanto's seeds are expensive and lose vigour after one generation, prompting the Indian Council of Agricultural Research to develop a cheaper Bt cotton variety with seeds that could be reused. The cotton incorporated the cry1Ac gene from the soil bacterium Bacillus thuringiensis (Bt), making the cotton toxic to bollworms. In parts of India cases of acquired resistance against Bt cotton have occurred.

The state of Maharashtra banned the sale and distribution of Bt cotton in 2012, to promote local Indian seeds, which demand less water, fertilizers and pesticide input, but lifted the ban in 2013.

India approved Bt cotton in 2002; now it accounts for 92% of all Indian cotton.

India approved Bt cotton in 2002; now it accounts for 92% of all Indian cotton. Average nationwide cotton yields went from 302 kg/ha in the 2002/3 season to a projected 481 kg/ha in 2011/12 up 59.3% overall. This chart shows the trends in yields, which took off after Bt was introduced in 2002. The graphs also show that and here comes ugly fact— in the last 4 years, as Bt has risen from 67% to 92% of India’s cotton, yields have dropped steadily.

BIOTECHNOLOGY IN MEDICINE

BIOTECHNOLOGY IN MEDICINE Genetically Insulin (Humulin) Engineered Insulin is a peptide hormone p roduced b y

Genetically

Insulin (Humulin)

Engineered

Insulin is a peptide hormone produced by beta cells in the pancreas of various organisms including human beings. It regulates the metabolism of carbohydrates an d fats by promoting the absorption of glucose from the blood to skeletal muscles and fat tissue and by causing fat to be stored rather than used for energy. Insulin also inhibits the production of glucose by the liver.

Structure:

Insulin is composed of two different types of peptide chains. Chain A has 21 amino acids and Chain B has 30 amino acids. Both chains contain alpha helices but no beta strands. There are 3 conserved disulfide bridges which help keep the two chains together.

disulfide bridges which help keep the two chains together. Need of Genetically Engineered Insulin: The original

Need of Genetically Engineered Insulin:

The original form of the wonder cure for diabetes, these were once the only type of insulin available, but are now rarely used. Animal insulin was originally made from ground-up animal healthy animals (slaughtered pigs & cows).

from ground-up animal healthy animals (slaughtered pigs & cows). pancreas tissue, and then later was extracted

pancreas tissue, and then later was extracted from

One of the problems with animal insulin was antibody issues. The body identifies them and tries to reject them.

Humulin:

Biosynthetic "human" insulin is now manufactured for widespread clinical use using genetic engineering techniques using recombinant DNA technology, which the manufacturers claim reduces the presence of many impurities, although there is no clinical evidence to substantiate this claim. Eli Lilly marketed the first artificial insulin, Humulin, in 1982.

Humulin production method is as follows:

1. DNA coding for A and B polypeptide chains of insulin are chemically synthesised a in the lab. Sixty three nucleotides are sequenced to produce A chain of insulin and ninety nucleotide long DNA designed to produce B chain of insulin, plus terminator codon is added at the end of each chain sequence. Anti-codon for methionine is added at the beginning of the sequence to distinguish humulin from the other bacterial proteins.

2. Chemically synthesized A and B chain DNA sequence are inserted into one of the marker gene which are present in the plasmid vector. Genes are inserted into the plasmid with the help of enzymes known as endonuclease and ligase.

3. The vector plasmids with the insulin gene are then introduced into the E. coli bacterial cell. These cells are then allowed to replicate by mitosis, along with the bacterial cell recombinant plasmid also gets replicated producing the human insulin.

4. A and B polypeptide chains of insulin are then extracted and purified from the fomenters in the lab. High-Performance Liquid Chromatography (HPLC) is used to get 100% pure humulin from the mixture of proteins.

5. The A and B polypeptide chains of insulin are mixed together and connected with each other by disulphide bond, forming the Humulin or synthetic human insulin.

Advantages & Disadvantages of Humulin:

Advantages & Disadvantages of Humulin: Humulin is the one and only human protein produced in the

Humulin is the one and only human protein produced in the bacteria with identical chemical structure to that of the natural human insulin. Administration of humulin reduces the possibility of antibody production and inflammatory response in diabetic patients. Major difficulty is the extraction of humulin from a mixture of host proteins present in the fermentation broth.

Now days to overcome this extraction problem synthetic human insulin are produced in the yeast cell instead of E. coli using the same procedure. As yeast is Eukaryotes they secrete the whole humulin molecule with perfect three dimensional structures, reducing the need for complex and time consuming purification methods. Now most of the diabetic patients are treated with synthetic human insulin. Small group of patients claim that episodes of hyperglycaemic complications have been increased after shifting from animal origin insulin to humulin. No study till date shows the difference between the frequency of hyperglycaemic complications in patient using humulin (synthetic human insulin) and animal origin insulin.

Gene Therapy

Gene therapy is the therapeutic delivery of nucleic acid polymers into a patient's cells as a drug to treat disease. Gene therapy is an experimental technique that uses genes to treat or prevent disease. In the future, this technique may allow doctors to treat a disorder by inserting a gene into a patient’s cells instead of using drugs or surgery. Researchers are testing several approaches to gene therapy, including:

are testing several approaches to gene therapy, including: • Replacing a mutated gene that causes disease

Replacing a mutated gene that causes disease with a healthy copy of the gene.

Inactivating, or “knocking out,” a mutated gene that is functioning improperly.

Introducing a new gene into the body to help fight a disease.

Although gene therapy is a promising treatment option for a number of diseases (including inherited disorders, some types of cancer, and certain viral infections), the technique remains risky and is still under study to make sure that it will be safe and effective. Gene therapy is currently only being tested for the treatment of diseases that have no other cures. It should be noted that not all medical procedures that introduce alterations to a patient's genetic makeup can be considered gene therapy. Bone marrow transplantation, and organ transplants in general have been found to introduce foreign DNA into patients. Gene therapy is defined by the precision of the procedure and the intention of direct therapeutic effects.

Gene therapy was conceptualized in 1972, by authors who urged caution before commencing human gene therapy studies.

The first attempt, albeit an unsuccessful one, at gene therapy (as well as the first case of medical transfer of foreign genes into humans not counting organ transplantation) was performed by Martin Cline on 10 July 1980. Cline claimed that one of the genes in his patients was active six months later, though he never published this data or had it verified and even if he is correct, it's unlikely it produced any significant beneficial effects treating beta-thalassemia.

The first germ line gene therapy consisted of producing a genetically engineered embryo in October 1996. The baby was born on July 21, 1997 and was produced by taking a donor's egg with healthy mitochondria, removing its nuclear DNA and filling it with the nuclear DNA of the biological mother - a procedure known as cytoplasmic transfer.

This procedure was referred to sensationally and somewhat inaccurately in the media as a "three parent baby", though mtDNA is not the primary human genome and has little effect on an organism's individual characteristics beyond powering their cells.

Gene therapy is a way to fix a genetic problem at its source. The polymers are either expressed as proteins, interfere with protein expression, or possibly correct genetic mutations.

The most common form uses DNA that encodes a functional, therapeutic gene to replace a mutated gene. The polymer molecule is packaged within a "vector", which carries the molecule inside cells.

The first commercial gene therapy, Gendicine, was approved in China in 2003 for the treatment of certain cancers. In 2011 Neovasculgen was registered in Russia as the first-in-class gene-therapy drug for treatment of peripheral artery disease,

including critical limb ischemia. In 2012 Glybera, a treatment for a rare inherited disorder, became the first treatment to be approved for clinical use in either Europe or the United States after its endorsement by the European Commission.

ADA deficiency is one form of SCID (severe combined immunodeficiency), a disorder that affects the immune system. ADA deficiency is very rare, but very dangerous, because a malfunctioning immune system leaves the body open to infection from bacteria and viruses.

leaves the body open to infection from bacteria and viruses. The disease is caused by a

The disease is caused by a mutation in a gene on chromosome 20. ADA deficiency is inherited in an autosomal recessive manner. The gene codes for the enzyme adenosine deaminase (ADA). Without this enzyme, the body is unable to break down a toxic substance called deoxyadenosine. The toxin builds up and destroys infection- fighting immune cells called T and B lymphocytes. Because ADA deficiency affects the immune system, people who have the disorder are more susceptible to all kinds of infections, particularly those of the skin, respiratory system, and gastrointestinal tract. They may also be shorter than normal. Sadly, most babies who are born with the disorder die within a few months.

Treatments of ADA Deficiency includes:

ADA enzyme in PEG vehicle

On September 14, 1990, the first gene therapy to combat this disease was performed by Dr. William French Anderson on a four-yearold girl, Ashanti DeSilva, at the National Institutes of Health, Bethesda, Maryland, U.S.A.

CONCLUSION

Biotechnology is the new wonder of science. It is truly multidisciplinary in nature and it encompasses several disciplines of basic sciences and engineering. The Science disciplines from which biotechnology draws heavily are microbiology, chemistry, biochemistry, genetics, molecular biology, immunology, cell and tissue culture and physiology. On the engineering side it leans heavily on process chemical and biochemical engineering since large scale cultivation of microorganisms and cells, their downstream processing are based on them. It comes to us as a great blessing

Biotechnology utilizes the technique called genetic engineering or recombinant DNA technology where a microorganism is isolated; its genetic material is cut, manipulated, sealed, again inserted in an organism and allowed to grow in a suitable environment under controlled conditions to get the desired product. It looks easy but is a very tedious job and it takes years for a research to achieve its goal. Like every other thing, biotechnology too has some harmful impacts:

1. Genetic engineering is a very vital part of biotechnology and the cost of transferring genes from one species to another is very expensive, which requires a huge amount of capital investment. The cost of producing genetically- modified plants and animals are sky- rocketing and the duration of return are also not predictable.

2. Genetic engineering crosses boundaries of reproduction by crossing genes of species that are completely unrelated; hence giving rise to hazardous results as well as also increasing the risk of harming multiple species.

3. When genetic material from certain viruses is used in the production of transgenic crops, there are chances that these virus genes will combine with crop genes to produce more destructive viruses. The consumption of such crops is hazardous to human health and can cause several life- threatening ailments. It can also result in cancer, often malignant as well.

4. Biotechnology also poses a number of environmental threats. Genetically modifies crops often infect monarch butteries and other insect species.

The applications of biotechnology are so broad, and the advantages so compelling, that virtually every industry is using this technology. Developments are underway in areas as diverse as pharmaceuticals, diagnostics, textiles, aquaculture, forestry, chemicals, household products, environmental cleanup, food processing and forensics to name a few. Biotechnology must continue to be carefully regulated so that the maximum benefits are received with the least risk.

Bibliography

http://www.genewatch.org/sub-568238

http://www.biotecharticles.com/Others-Article/Human-Insulin-and-

Recombinant-DNA-Technology-70.html

http://www.diabetes.co.uk/insulin/animal-insulin.htmlBiology textbook (N.C.E.R.T) Class 12 th