cena 12 BIOTECHNOLOGY AND ITS APPLICATIONw Biology CONTENTS BIOTECHNOLOGICAL APPLICATIONS IN ‘AGRICULTURE BIOTECHNOLOGICAL APPLICATIONS 1 ‘MEDICINE TRANSGENIC ANIMALS AND ETHICAL “It always seems impossible until its done”. — Nelson MandelaBiotechnology and its Application vw 12-1 BIOTECHNOLOGICAL APPLICATIONS IN AGRICULTURE Biotechnology offers a wide range of applications, including biopharmaceuticals, therapeutics, diagnostics, agriculture based genetically modified crops, processed foods, bioremediation, waste management and energy generation '¢ The following are 3 major biotechnology research areas: i. Providing the most effective catalyst in the form of a better organism, such as a microbe or pure enzyme. li, Using engineering to create ideal conditions for a catalyst to act, and iii, Purification of the protein/organic compound using downstream processing technologies. ‘¢ The three approaches for improving food production that can be considered are: Agriculture based on ‘agrochemicals Organi agriculture Use of agriculture based on genetically engineered crops ee ‘+ The Green Revolution was an era in India's history when modem methods and technology, such as the usage of high yielding variety (HYV) seeds, was used to transform agriculture into an industrial system. Green revolution was led by M.S. Swaminathan. ‘+ The Green Revolution tripled food production, yet it was stil insufficient to sustain the growing population. Improved crop types have contributed to increased yields, but better management methods and the use of agrochemicals (fertilisers and pesticides) have played a greater role, The usage of genetically modified crops helps to minimise the need of fertilisers and pesticides, which had negative environmental consequences. 12.1.1 GENETICALLY MODIFIED ORGANISMS (GMO) '* Plants, bacteria, fungi, and animals, that have their genetic material manipulated are known as genetically modified organisms (GMOs) or transgenic species/plants. 83Biology ‘¢ Genetic modification has the following effects: (i) Crops that are more resistant to abiotic stresses (cold, drought, salt, heat) (ii) Chemical pesticides (pest-resistant crops) are used less frequently (ii) Helped in the reduction of post-harvest losses (iv) Higher efficiency of mineral use by plants (this avoids soil fertility from being exhausted) (v) Production of food with a higher nutritional value, such as golden rice, which is vitamin ‘A’ enriched rice. (NET 2019) (vi) Create custom-made plants to supply industry with alternative resources (starch, fuel, medications, and so on) Pest Resistant Plants ‘¢ They act as bio-pesticide. ‘¢- Itreduces the need for insecticides. + Bt cotton, Bt com, rice, tomato, potato, soyabean, and other genetically modified crops are some examples. Bt Cotton ‘Some Bacillus thuringiensis (Bt) strains produce proteins that kill insects such as beetles, lepidopterans (tobacco budworm, armyworm) and dipterans (flies, mosquitoes). (NEET 2020) ‘* During a specific phase of their growth, B. thuringiensis produces @ harmful insecti protein (Bt toxin). Because it exists as inactive protoxins, it does not kill the Bacillus. ‘+ The insect consumes this Bt toxin ( inactive form of toxin ). As this toxin reaches the gut of insect due to the alkaline pH , the toxin gets converted into an active form. (Mains 2011). This, alkaline pH also solubulizes the crystals of Bt toxin. (NEET 2019) 84]Biotechnology and its Application vw = ‘¢ The toxin attaches to the surface of epithelial cells in the midgut, thus creating pores. It causes the insect's cells to swell, undergo lysis, and finally death of insects, ‘* Bt toxin genes were obtained from B. thuringiensis and inserted into many crop like cotton plants, ‘¢ The majority of Bt toxins are insect-specific. Cry is the name of the gene that codes for the toxin. Proteins produced by the genes cry/Ac and cryllAb, for example, control cotton bollworms. Proteins of cryiAb gene also control the com borer. Eau a Protoxin Basic an pH tiv Bt toxin erystal ae Ee” Solubilisation Activation Insect Bind to receptor 2 -o Cia) cocoa Pore formation so -~A-A Increase permeability Fig 12.4.4 Action of Bt toxin crystal on insects Nematode Resistance in Tobacco Plants ‘¢ Meloidegyne incognitia, a nematode, infects the roots of tobacco plants (NEET-I 2016), reducing yield, ‘¢ RNA interference (RNAi) is a strategy that can be used to prevent infestation caused by this nematode. ‘¢ RNAiisa cellular defence mechanism found in all eukaryotes. Due to a complementary dsRNA. molecule, it inhibits translation of a specific mRNA (silencing). (NEET 2019) ‘¢ Gene silencing mechanism involves dsRNA which when introduced inside pest can lead to death of the pest (nematode) ‘¢ This complementary RNA comes from an infection by RNA viruses or mobile genetic elements (transposons) that replicate with the use of an RNA intermediate. ‘+ Nematode-specific genes (DNA) are transferred into host plants using Agrobacterium vectors, In host cells, it creates both sense and anti-sense RNA (Mains 2012). These RNAs are complementary. As a result, double-stranded (ds) RNA is formed. 85‘It initiates RNAi and silences the nematode’s specific mRNA. As a result, the parasite is tunable to thrive in a transgenic host that produces specific interfering RNA. Inecreo pant Agrobacetur ranser gion a Fig. 12.1.2 Nematode-specific genes transferred Fig, 12.1.3 Tabacco root infected by nematode into host plant using Agrobacterium vector (Root knot nematode) te 2 BIOTECHNOLOGICAL APPLICATIONS IN MEDICINE The recombinant DNA technique aids in the mass manufacture of more effective and safe therapeutic medications. '¢ Unwanted immunological responses are induced by products derived from non-human sources. Recombinant therapeutics, on the other hand, do not have these issues, '* Approximately 30 recombinant therapeutics have been approved till date. In India, 12 of these are being sold, 12.2.1 GENETICALLY ENGINEERED INSULIN ‘* Adult-onset diabetes can be controlled by taking insulin at regular intervals. ‘¢ Insulin produced by the pancreas of animals (cattle and pigs) generates .) allergic or other reactions to the foreign protein. '¢ Its now possible to make human insulin from bacteria 86Biotechnology and its Application ‘+ Insulin is made up of two short polypeptide chains (chain A and chain B). Disulphide bridges connect chains A and B. (NEET-I 2016) ‘¢ Insulin is produced as a pro-hormone (pro-insulin) in mammals. It must be processed in order to produce a fully functional hormone, ¢ € peptide is an additional stretch found in the pro-hormone (NEET 2020). During the ‘maturation of insulin, this is eliminated. + In 1983, Eli Lilly (an American company) developed two DNA sequences that corresponded to the A and B chains (peptide) of human insulin and used them to manufacture insulin chains in E. coli plasmids (NEET 2020), To make human insulin, disulphide connections were formed between chains A and B. Proinsulin 90000000006 poo C-peptide POD, & crane CCUTATC Tatts eset 1: OBCOOOPODEDESDOOOO EHO om Insulin f NI COOH Behain ‘1 23 45 67 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 25 24 25 26 27 28 29:30 Fig. 12.2.4 Proinsulin maturing inte insulin | 87w Biok 12.2.2 GENE THERAPY ‘¢ Gene therapy is a set of techniques for correcting a gene defect that has been identified in a child or embryo. ¢ To treat a hereditary condition, genes _ are injected into a person's cells and ace tissues. It compensates for the gene that is not functional ¢ A 4-year-old girl child with adenosine deaminase (ADA) deficiency received the first clinical gene therapy in 1990 (NEET-II 2016). The condition is caused by the deletion of the adenosine deaminase gene (an enzyme required for the proper functioning of the immune system). Bone marrow transplantation or enzyme replacement therapy (injection of functional ADA) can be used to cure it (AIPMT 2009). However, these methods are not completely curative. ‘¢ Therefore, ADA-SCID gene therapy (ex vivo gene therapy) was given, which may act as effective cure steps of this gene therapy given in figure 12.2.2. ‘¢ Asa first step towards gene therapy, lymphocytes from the blood of the patient are cultivated in a culture in vitro. ‘¢ Afunctional ADA cDNA (through a retroviral vector) is then inserted into these lymphocytes, after which they are returned to the patient. ‘* However, because these cells are not immortal, the patient will need to get infusions of genetically altered Iymphooytes on a regular basis. ‘¢ However, a permanent cure could be achieved if gene isolate from marrow cells that produce ADA, is injected into cells at an early embryonic stage. a I-— ») os SB ~— Fe? Fig, 12.2.2ADA-SCID gene therapy 88Biotechnology and its Application 12.2.3 MOLECULAR DIAGNOSIS ‘¢ tis impossible to detect and diagnose diseases early using conventional methods (serum and urine analysis) ‘¢ Early diagnosis approaches include Recombinant DNA technology, Polymerase Chain reaction (PCR), and Enzyme yey GLa \ked Immuno-Sorbent Assay (ELISA). Polymerase Chain reaction (PCR) ‘¢ Symptoms alone are usually enough to suspect the presence of a disease. By this point, the pathogen’s concentration in the body is extremely high ‘¢ Amplification of their nucleic acid by PCR, on the other hand, can identify very low concentrations of bacteria or viruses, Uses of PCR Fig, 12.2.3 PCR Machine ‘¢ To identify HIV in AIDS patients who are suspected of having the disease. ‘¢ To look for gene mutations in cancer patients who have been diagnosed with the disease. ‘¢ Tofind out about a variety of other genetic disorders. In any clone of cells, a single stranded DNA or RNA tagged with a radioactive molecule (probe) which is allowed to hybridise to its corresponding DNA. This is followed by autoradiography based detection. Because the probe does not have complementarity with the mutant gene, the clone with the mutated gene will not appear on the photographic film. Enzyme Linked Immuno-Sorbent Assay (ELISA) ‘¢ Itis based on the principle of antigen-antibody interaction. ‘¢ The presence of antigens (proteins, glycoproteins, etc.) orantibodies generated against the pathogen can be used to detect pathogen infection Fig. 122.4 ELSA |s9Biology z 3 TRANSGENIC ANIMALS AND ETHICAL ISSUES Transgenic animals are animals that have had their DNA manipulated to possess and express an additional (foreign) gene. ‘* Transgenic rats, rabbits, pigs, sheep, cows, and fish have been created using transge! method with trans genic mice accounting for approximately 95% of all transgenic species. Benefits of Transgenic Animals Normal physiology and development ‘¢ The objective is to learn more about gene regulation and how it affects the body's regular functioning and development. ‘¢ For example, study on complicated factors like insulin growth factor. The biological effects of genes (from other species) that influence the production of this factor are studied. This provides details on the factor's biological function in the body. Study of disease ‘+ Transgenic animals are being used to study the role of genes in the development of disease and, their, new treatments. * For example, transgenic models for cancer, cystic fibrosis, rheumatoid arthritis and Alzheimer's disease. Biological products ‘© Biological products are used in medicines to cure specific human ‘ diseases, although they are often expensive to produce. aS ‘+The introduction of the segment of DNA (or genes) that codes for a Py, specific product can result in. prod Useful biological products. ‘* For example, human protein (a-t-antitrypsin) used to cure emphysema and products for treatment of phenylketonuria (PKU) . and eystie fibrosis ete.technology and its Application Rosie was the first transgenic cow which produced milk (2.4 gm per litre) that was enriched with human protein. Itis a nutritionally more balanced product for human babies than natural cow’s milk since it contains human a-lactalbumin. Vaccine safety ‘¢ Transgenic mice are being developed to test vaccines for safety before they are used on humans. (AIPMT 2010) ‘*- Ifit proves to be reliable, they may be able to replace the use of monkeys in vaccine testing. ‘+ For example, polio vaccine is being tested using transgenic mice. & ¥ Vaccine production is the most common application of animal cell culture technology today. (AIPMT 2003) Chemical safety testing (toxicity testing) '¢ Transgenic animals are produced with genes that increase their sensi compared to non-transgenic animals, to toxic substances ¢ They are subjected to the toxic substances and observed consequences. It produces immediate outcomes. 12.3.1 ETHICAL ISSUES '¢ Without regulation, the human race will continue to exploit biological organi activity that may help or hurt living beings must meet some ethical standards. ‘¢ Unpredictable outcomes may arise from genetic modification (GM). ‘* The Indian government has established organisations such as the GEAC (Gené isms, All human Engineering Approval Committee) to decide on the validity of genetic engineering research and the safety of GM organisms while introducing them for public services. (NET 2018) \91w Biology Patent Issues ‘¢ There is growing public outrage that certain companies are being granted patents for products and technology that use genetic materials, plants, and other biological resources that have been identified, produced, and used by farmers and indigenous people in a particular region or country. For example: Basmati rice, herbal medicines (turmeric, neem etc) In India, there are an estimated 200,000 rice types. Basmati rice has a unique scent and flavour. Basmati exists in 27 different types in India. The US Patent and Trademark Office granted an American corporation £ patent rights on Basmati rice in 1997. This allowed the company to sell a ‘new’ Basmati type (NEET 2018). This was derived from the varieties of Indian farmers. ‘Anovelty was created by crossing Indian Basmati with semi-dwarf varieties. The marketing of Basmati rice by others may be restricted by a patent. Biopiracy ‘It refers to the use of bio-resources by multinational corporations and other organisations without legal permission from the countries and people concered, and without payment of, compensation. (NEET 2018) ¢ The majority of developed countries are wealthy financially, yet they lack biodiversity and traditional wisdom. The developing and undeveloped countries, on the other hand, are rich in biodiversity and traditional bio-resource knowledge. '* Traditional bio-resources knowledge can be put to use in the development of current biotechnology based applications. 92Biotechnology and its Application = ‘+ Therefore it is important to develop law for preventing unauthorised exploitation of bio- resources and traditional knowledge. ‘¢ The Indian Parliament has approved the second amendment to the Indian Patents Bill, which addresses concerns such as patent terms, emergency measures, and research and development initiatives. TCM ee TRAIN YOUR BRAIN Question ‘A few companies in China and South Korea offer dog cloning services, using cells from their customer's pet. Should their clients anticipate the clone to resemble their original pet in appearance? Why do you think that is? What ethical issues does this raise? Solution : No, primarily because the clone's surroundings differ from that of the original pet in subtle (and possibly not so subtle) ways. This raises certain ethical concerns. Several hundred embryos were cloned to become Dolly (sheep), a mammal, but only one lived to adulthood. 93