Sample Materials for CSAT Paper -1 (G.S.) Pre 2013: "Science & Technology: Biotechnology"

Subject: Science & Technology:
Topic: Biotechnology

Meaning, Scope and Organisational Set Up

A. Concept

Biotechnology is the use of complete living cells or part of living cells to produce new or improved products of service systems. According to the U.S. National Science Foundation Biotechnology consists of "the controlled use of biological agents, such as, micro-organisms or cellular components, for beneficial use." The European Federation of Biotechnology define "Biotechnology as the integrated use of biochemistry, microbiology, molecular biology and engineering sciences in order to achieve technological application of the capabilities of micro-organisms, cultured tissues/cells and part thereof”. In the modern context, an illustration of biotechnology would be the practice, for centuries, of fermentation of wine into alcohol using micro-organisms.

B. Background

The crucial advance in biotechnology that laid the foundation took place in 1972 when the first successful venture namely direct insertion of foreign DNA, the genetic material, in a host organism was carried out in the U.S.A. The technique was termed recombinant DNA (rDNA) technique. This in effect opened up immense possibilities of direct manipulation of the genetic material, the blueprint of all living organisms, to produce chemicals and other products needed by man. This pioneering work resulted in the award of the Noble prize to Professor Paul Berg of Stanford University and others. The rDNA technique along with "cell fusion" -or "hybridoma" technology constitutes broadly the areas of modern biotechnology. The cell fusion technique was developed in 1971 by Dr. Milsein Kohler and Jeme. As a reaction to an antigen the host organism produces what are called antibodies. This is basically a defence mechanism. The cell fusion technique has immense potentialities for specific diseases arising out of viral, bacterial and other microbial infection. A distinction is made between 'non-gene biotechnology' and 'gene biotechnology': the former works with whole cells, tissues or even individual organisms; the latter deals with transfer of genes from one organism to another or genetic engineering. Non-gene biotechnology is a more popular practice, and plant tissue culture, hybrid seed production, microbial fermentation, production of hybridoma antibodies are widespread biotechnology practices in our Country.

C. Organisation And Manpower

In 1982, Government of India setup the National Biotechnology Board which was replaced by a separate Department of Biotechnology (DBT) in the Ministry of Science and Technology in 1986. This was in recognition of the need for a focal point in the administration and structure of the government for planning promotion and co-ordination of biotechnological programme. The main responsibilities of the Department of biotechnology are:
i) To evolve integrated plans and programmes in biotechnology;
ii) To identify specific R&D programmes in biotechnology and biotechnology related manufacturing;
iii) Establishment of infrastructure support at the national level;
iv ) To act as an agent of the Government for import of new recombinant DNA based biotechnological processes, products and technology;
v) To evolve bio-safety guidelines for laboratory research production and applications;
vi) To initiate scientific and technical, efforts related to biotechnology;
vii) Programmes of manpower development in the areas of biotechnology;
viii) Establishment of international centre for genetic engineering and biotechnology,

D. Institutions

1. Centre for DNA Fingerprinting and Diagnostics (CDFD), Hyderabad: CDFD provides service for DNA fingerprinting, diagnostics, bioinformatics and automated genome analysis and undertakes research and developments in the area of genetics, molecular and cellular biology, molecular pathogenesis and bioinformatics.

2. Institute of Bio Resources and Sustainable Development (IBSD), Imphal (Manipur): IBSD has been established with to develop utillse the rich bio-resources of the North-Eastern region of the country through the application of modern tools of biology and biotechnology. The institute works on medicinal, horticultural, microbial, insect and aquatic resources as well as eco restoration.

3. Institute of Life Science (IIS), Bhubaneshwar: The Institute of life sciences has been established to conduct basic and applied research in frontiers areas of life sciences, to promote interaction among scientists, conduct inter-disciplinary research and for dissemination of scientific knowledge.

4. National Institute of Immunology (NII), New Delhi: NII's primary responsibility is to help create the scientific base for innovations of relevance for the development of the nation. Its main areas of research are infection and immunity, molecular design, gene regulation and reproduction and development.

5. National Centre for Plant Genome Research (NCPGR), JNU, New Delhi: NCPGR works in core research area of plant genomics covering structural, functional and application components of genomics. The institute has been working on nutritional genomics of potato, structural and functional genomics of chickpea.

6. National Brain Research Centre (NBRC), Gurgaon: NBRC undertakes basic research to understand brain function in diseases and normal conditions. NBRC has established networking centres with 35 foreign institutions like NIMH, USA for research co-operation and training.

7. National Centre for Cell Sciences (NCCS), Pune: NCCS undertakes research and development at the cutting edge of cell sciences, teaching, training in addition to providing services as a national repository for cell lines and hybridomas. NCCS has developed technologies for preservation and revival of bone marrow stem cells which led to a successful transplantation in a neuroblastoma patient.

8. International Centre for Genetic Engineering and Biotechnology (ICGEB), New Delhi: ICGEB focuses on basic research in human diseases and agriculture, especially to look at problems of the Indian subcontinent. IGCEB has 'state-of-art' facilities and it will be a National facility to be used by scientific community. It has developed technology for hepatitis C disgnostic kit and a malaria vaccine program is progressing as per time Schedule.

9. Bharat Immunological and Biological Corporation Limited (BIBCOL), Bulandshahr: BIBCOL, a public sector undertaking has the most modern manufacturing unit based on good manufacturing practice requirements as specified by WHO and US Federal standards.

10. Biotechnology parks and incubators: The Biotechnology Parks and Biotech Incubation Centres established provide a good template for the promotion of Biotech startup companies and the promotion of Public Private Partnerships. Biotech Park and incubation Centres have been established at Lucknow, UP and Shapoorji Pallonji Biotech Park, Genome Valley, Hyderabad (AP). The other projects approved for Himachal Pradesh, Karnataka and Kerala for setting up of biotech incubation/pilot plant facilities are at various stages of development.

(E) National Bio Resource Development Board (NBDB)

Following the Finance Minister's Budget Speech, 1999, a National Bio Resource Development Board (NBDB) has been set up under the chairmanship of Minister of Science and Technology. NBDB shall decide the broad policy framework for effective application of biotechnological and related scientific approaches for R&D and sustainable utilisation of bio-resources, especially for development of new products and processes. It shall develop a scientific plan of action for contribution to the economic prosperity of the nation through accelerated research and development using the modern tools of biosciences. The NBDB shall adopt both resource based and region- based approaches. It shall also be involved in training, capacity building and awareness generation in bio-resources.

A National steering committee has been constituted to support the activities of the board; the board has identified 3 priorities:
1. Preparation of digitised inventories of plant, animal, microbial and marine resources.
2. R&D projects, programmes support, establishment of centres of excellence, training activities and demonstrations, for the development of bio resources of special areas such as North-eastern region, Himalayan region, coastal and land ecosystems, desert region etc.
3. Knowledge empowerment and human resource training would be a priority for the board.

Ques. 1 : Give an account of the techniques of Biotechnology?

Ans. The main techniques of biotechnology are - genetic engineering, cell culture, tissue culture, bio-processing, protein engineering, monoclonal antibody production and biosensor technology. As has been recognised all over the world, in the last fifteen years, there has been revolution in the field of Biotechnology as evidenced through new discoveries and inventions in the areas of isolation and manipulation of genes, better understanding of biological molecules, the advent of recombinant DNA technique enabling the genes to be transferred between organisms to produce scare proteins of plant and animal origin as also human growth factors and hormones.

Ques. 2 : Discuss in brief the Genetic engineering?

Ans. Genetic Engineering : The utilisation of genetic machinery of life for production of any special substance is called gene technology or genetic engineering. The genetic modification of micro- organisms, so vital for their utilisation in the production of useful biochemical, can be brought about by simple recombination or by complex genetic manipulations. Some of the techniques are:
Isolation of Genes: Appropriate sequence of genes is directly obtained from genome of normal cell or from other cells. This is made possible by cleavage and denaturation of DNA extracted from the cells.
Synthesis of Genes: This is done by chemical methods. Dr. Hargobind Khurana reported this in 1970.
Recombinant DNA: Breakage of DNA molecule at two desired places to isolate a specific DNA fragment and then inserting it in another DNA molecule at a desired position results in a new gene product which is called as recombinant DNA (r-DNA). The receiving organism is said to be transgenic. Using this technique we can isolate and clone single copy of a gene or a DNA molecule into an indefinite number of copies, all identical.
Gene Cloning: Isolation of gene and replication of a single copy of gene or DNA segment into an infinite number of copies, all identical, is known as gene cloning. This becomes possible because vectors like plasmids and phages reproduce in their usual style even after insertion of foreign DNA. This inserted DNA will also replicate faithfully with parent DNA. Recently extensive use of newly discovered polymerase chain reaction (PCR) has also been made for gene technology.

Ques. 3 : What is Tissue Culture?

Ans. Tissue Culture : Tissue culture is the technology of artificially growing micro-organisms or cells or tissues or organs to the desired genetic purity with properties such as high yield and disease resistance.

The microbes in culture are used in recombinant DNA technology and in a variety of industrial processes, plant cells and tissues are used for a variety of genetic manipulations. For example another culture is used for haploid breeding; gametic and somatic cell/tissue cultures are used for tapping gametoclonal and somaclonal variations or for production of artificial seeds. Transformation of protoplast in culture leads to production of useful transgenic plants. Embryo culture technique has also helped extending the range of distant hybridisation for plant breeding purposes. Animal cells are used for multiplication of superior livestock using a variety of techniques like Cloning of superior embryonic cells, transformation of cultured cells leading to the production of transgenic animals and in vitro fertilisation and transfer of embryos to surrogate mothers.

Ques. 4 : Briefly discuss microbial bio-technology?

Ans. Microbial Biotechnology : Micro organisms have been harnessed by man for the production of useful materials. The latest initiatives in Microbial Biotechnology have been taken in the following matters:
i) Rehabilitation of degraded land such as alkaline soil, mine dumps and dump from metallurgical factories, utilising microbial supports.
ii) Dissolution of pyretic shells by microbial methods to liberate entrapped noble metals like gold, silver etc. through the process of bio-techning of low and lean grade orders.
iii) Degradation of polyphenolic compounds using microbial approaches.
iv) Standardisation of shuttling vectors for E-coli and streptomyces having capabilities of accepting chester genes of Ansamycines.
v) Strengthening of microbial teaching and research in identified universities.
vi) Development of fungicides to contain fungal infections in plants and vegetables.
vii) Development of microbial enzymes active in extreme temperatures, novel antibiotics and bioactive proteins and other bio-molecules for industrial use.

Ques. 5 : Briefly discuss the concept of Human Genetics?

Ans. Human Genetics : Genetic diseases have posed a serious threat to the health in the Indian population. This has led to an increased demand for genetic counselling and screening tests both for carrier detection and for identifying pregnancy at risk. Currently prenatal diagnosis is possible for most chromosomal disorder and many major congenital malformations.
Diagnostic services are being provided to Thalassemia affected families and prenatal diagnosis of pregnant women in those families. In AIIMS, New Delhi investigations relating to haemophilia are carried out in various families affected by the disease and prenatal diagnosis had helped in identifying the foetus that carries the disease in pregnant women in those families. In North-east, Sickle-Cell anaemia was traced in certain tribal populations through screening of blood samples for Haemoglobin-E in those persons. Based on the haematological results, it has been possible to construct a linear discriminate function to identify suffering individuals with an error probability of less than 5%.

Formation of gene is a very complex issue. Scientists have not achieved full control over their formation. Indian Scientists have developed good expertise to understand gene structure and therefore, it would be possible to identify genetic defects.

Genes consists of DNA. Each gene has a different sequence of bases. Each sequence has coded information that ultimately leads to the production of a specific protein. These molecules govern several life processes. Specific clinical trials are under way in case of a few selected genetic disorders. Gene therapy is being tried only in terminal cases or where there is no other avenue for survival. It is hoped that gene therapy will become a serious option in about 5 to 10 years.

Ques. 6 : What is DNA fingerprinting? Discuss its importance?

Ans. DNA Fingerprinting : DNA finger printing technique was first developed by ALEC JEFFREYS in 1985- 86 in UK. DNA finger printing is a technique, by which an individual can be identified at molecular level; the technique identifies the repeating sequences in the DNA that are unique to a particular individual.

DNA is the basic genetic material. It not only carries a blue print for our life, but also varies significantly from one person to another. What DNA fingerprinting does is to look inside DNA, regions of DNA that show a great deal of variations from one person to another. These regions account for a small proposition of our genetic material, but the variations are such that we can locate these regions, highlight and identify them using DNA probes and obtain a pattern, a series of bands or stripes on X-ray film. These DNA patterns / sequences are essentially unique to an individual, except in case of identical twins who have the same DNA.

The samples required for DNA fingerprinting examination are drop of blood, semen, saliva, and any body part such as bones, tissue, skull teeth, hair with root etc.
DNA fingerprinting has revolutionised forensic medicine and its applications now, encompass broad areas. The technology has made it possible to identify the source of biological samples at scenes of crime. This will resolve disputes of maternity paternity, identification of mutilated remains, identifications of rape/murder, identification of missing child, exchange of babies in hospital words, forensic wildlife, investigate family relationships in animals, protection of farmers rights and biodiversity (technology was used to prove the genetic distinctiveness of Indian Basmati rice composed to Pakistani variety).

Latent of fingerprint was discovered by Sir William Hesschel, a British revenue office in British India in 1860. Latent fingerprint is the identification of the individual by the impression of the fingers. While DNA fingerprinting is the identification of an individual, by the genetic markers, which are present on chromosomes. Latent fingerprints can be distorted by surgery. Latent fingerprint despite its limitations has come to be a handy tool in crime investigation over the last 140 years.
In India, DNA fingerprinting using B Km probe has been developed by Dr. Lalji Singh at Centre for Cell and Molecular Biology (CCMB), Hyderabad. DNA fingerprinting tests are performed at centre for DNA fingerprinting and diagnostics (CDFD). Hyderabad, a new DNA typing laboratory has been established Central Forensic Science Laboratory at Kolkata by the Bureau of Police Research and Development. Thus, the country has two DNA typing facilities.

Ques. 7 : What is Hybridoma?

Ans. Hybridoma : Hybridoma is a cell produced by" fusion of an antibody- producing cell and a mycloma cell (tumour of B-Iymphocyte). The technique of fusing mycloma cell with antibody-producing cell is called somatic cell hybridisation. Kohler and Milstein were awarded Nobel Prize in medicine in 1984 for the development of 'hybridoma'. The value of hybridomas was not appreciated until monoclonal antibodies were regularly produced in rodents for diagnostics.

Ques. 8 : What is Monolonal antibodies? Point out its uses?

Ans. Monoclonal antibodies are:
• Antibodies of exceptional purity and specificity
• Components of immune system
• Able to recognize and bind to a specific antigen.
Monoclonal antibodies (m Ab) are antibodies* that are identical because they are produced by one type of immune cell, all clones of a single parent cell.

Production

If a foreign substance (an antigen) is injected into a vertebrate such as a mouse or a human, some of the immune systems B-cells will turn into plasma cells and start to produce antibodies that bind to that antigen. Each B-cell produces only one kind of antibody, but different B-cells will produce structurally different antibodies that bind to different parts ("epitopes") of the antigen. This natural mixture of antibodies is known, as polyclonal antibodies.

To produce m Ab, B-cells from the spleen of an animals which have been challenged with antigen are removed. These b-cells are then fused myeloma cells (myeloma is a B-cell cancer/tumour). The fused hybrid cells (hybridomas), being cancer cells, multiply rapidly and produce large amounts of antibodies. From the hybridomas, we can obtain a number of different colonies but each colony produces only one type of antibody (hence MONOCLONAL). Among the various types of antibodies, the variety that effectively bind with antigen is then picked out.

Uses

Monoclonal antibodies are widely used as diagnostic and research reagents. They are currently utilized in many diagnostic procedures including:
• Measuring protein and drug levels in serum
• Typing tissue and blood
• Identifying infectious agents
• Identifying the specific cells involved in immune response
• Identifying tumour antigens and auto-antibodies.

Ques. 9 : What is Artificial Insemination?

Ans. Artificial Inseminations : Artificial Insemination (AI) is the artificial introduction of semen into the reproductive tract of female.
Depending upon the location of sperm insemination, Artificial Insemination is of different types:
a) Intra-cervical: semen placed into the cervical canal
b) Intra-uterine: semen inseminated inside the uterine cavity
c) Intra-follicular: semen introduced in the ovarian follicle
d) Intra tubal: semen placed in the fallopian tube.
Of the 4 different forms of Artificial Insemination, intrauterine insemination (IUI) is the most commonly used form.

Ques. 10 : Discuss in brief the Artificial Insemination in animals?

Ans. Artificial insemination in animals : Modern techniques for AI were first developed for the dairy cattle industry to allow many cows to .be impregnated with the-sperm of a bull for improved milk production, Now AI is used in various animals like horses, swine, pedigreed dogs, honey bees etc to propagate desirable characteristics of one male to many females or to overcome breeding problems.

In Artificial Insemination, the semen that is going to be inseminated is first collected, then frozen and later transported to the female's location. To allow the sperm to remain viable during the time before and after it is frozen, the semen is mixed in with a solution containing glycerol, in order to allow the semen from a donor to impregnate more female, an "extender" solution is added-to the donor semen so that insemination is possible with fewer sperms. Antibiotics such as streptomycin diseases, (sexually- transmitted diseases)

Ques. 11 : What do you understand by Human Artificial Insemination?

Ans. Human Artificial insemination : In humans, Artificial Insemination is usually part of an infertility treatment. The sperms are either of husband (artificial insemination by husband, AIH) or donor (artificial insemination by donor, AID).
In its simplest form, the women's menstrual cycle is closely observed and just when an ovum is released, semen from donor is place in women's reproductive tract. If the procedure is successful, she conceives and bears a baby, making her both the genetic and gestational mother.
Artificial Insemination is recommended when these are structural abnormalities in women, moderate male factor infertility, cervical mucus insufficiency, hostile cervical mucus etc.
Artificial Insemination is advantageous over invasive procedures like In Vitro fertilisation (IVF) as it is less invasive, relatively uncomplicated, and economical. Artificial Insemination however has resulted in debated revolving around surrogate parenting. Legal issues have arisen in cases where the gestational (and possibly genetic) mother decides to keep the child. Also, there have been debates over thoughts of sperm donors. Some also argue that conceiving a baby without sexual intercourse is not ethical.
Ques. 12 : Briefly discuss the In vitro fertilization?
Ans. In Vitro Fertilisation (Test Tube Baby): ‘In vitro’ in Latin means 'in glass' refers to the test tubes, But in 'in vitro' fertilisation neither glass or test tubes are being used, this term (in vitro) is used generally for lab procedures.
In Vitro Fertilization (IVF) is a major treatment in infertility when all other methods of achieving conception have failed. IVF is a technique in which egg cells are fertilized outside the woman's body.
In this technique, ova (eggs) are removed from woman's ovary and sperms are allowed to fertilize them in a fluid medium. The fertilized egg (zygote) is then transferred to the female's uterus to establish a successful pregnancy.
The first ever 'test-tube baby' Lousie Brown was born on July 25, 1978 by this technique. This technique was developed to overcome infertility due to problems of fallopian tube, but is now the most successful method to overcome infertility cost considerations are the major drawback of this technique.

Ques. 13 : Give an account of Embryo Transfer?

Ans. Embryo Transfer : Embryo transfer refers to a step in process of In Vitro Fertilization (IVF) where one or several embryos are placed into the uterus (womb) of female in order to establish a pregnancy. In IVF, fertilisation between egg and sperm occurs outside female body. Once, fertilization occurs, the resulting embryo has to be transferred back into womb for its development. This method of transfer of fertilized egg is called embryo transfer.

Embryo can be transferred as either "fresh" from fertilized egg cells of same menstrual cycle or "frozen" (i.e.) they have been generated in a preceding cycle and then cryopreserved. Before the embryo is implanted inside the womb it must be-ensured that the uterine wrong (endometrium) is appropriately prepared. The embryo's are generally transferred 3 days after fertilization. The procedure of embryo transfer is preferred with the aid of ultrasound to allow for precise placement.

The Science and Technology project on Embryo Transfer Technology being implemented in mission mode since 1987 had been successfully completed. The embryo recovery in cattle and buffalo and establishment of embryo transfer and related techniques had come close to the international standards. The four Regional Centres and 25 State Level Centres would provide the infrastructure for training and actual application/utilisation of this technology. A stock of superior germplasm has beer established to provide 100 embryo free of cost to the milk co-operative societies to benefits the farmers. A large number of scientists have been trained and the expertise has been developed in the areas of super ovulation, embryo splitting, cryopreservation etc. The technology packages generated under this project will be transferred to the National Dairy Development Mission and the National Bull Production Programme. Considerable progress was made on embryo sexing, splitting and in vitro fertilization and maturation other activities in the area of animal birth control vaccine, TALSUR; establishment of the genetically superior stock of cattle; improvement in the growth rate of poultry broilers, development of immuno-diagnostics are under implementation with promising results. The open nucleus breeding system (ONBS) for production of crossbreed Sahiwal Bulls in under progress and the first batch of bulls is expected soon.

Ques. 14 : What is Recombinant DNA technique? Point out its uses?

Ans. Recombinant DNA : Recombinant DNA is any DNA molecule that has been manipulated by in vitro procedures to create a novel sequence. The recombinant molecule produced may have a modified base sequence or contain sequences from two or more different genes or organisms. They are usually introduced into an organism to create a novel protein or R NA molecule that alters the properties of the organism.

Major recombinant DNA techniques include site-directed mutagenesis, doming and polymerase chain reaction (PCR). Key enzymes required for recombinant DNA work are type II restriction enzymes: DNA Ligase, DNA polymerase, reverse transcriptase and ON.A phosphatase.

Some of the uses of recombinant DNA techniques are:
• To done genes
• To knock out a gene in an organism
• To create: anti-sense RNAs in an organism so as to interfere with gene expression.
• To add a gene(s) to an organism to engineer a new or modified metabolic pathway.

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Ques. 15 : What is proteome?

Ans. Proteome : 'Proteome' means protein complement expressed by a genome, the proteome all the expressed genes or proteins of a genome., A cell type may display numerous sub proteomes under different growing conditions, nutrition status, health or disease "Proteomics" is the use of quantitative measurements of the level of a protein or gene expression to characterize biological processes to decipher the mechanism and control of gene expression. Proteome research / proteomics is the best path between genome and function studies.

Glossary of Biotech Terms

1. Antibody: is a protein secreted by B-Lymphocytes in response to an antigen.
2. Antigen: is a foreign substance that induces the formation of antibody.
3. Myeloma: is a tumour of B-lymphocyte cells arising in the bone marrow,
4. Bacteriophage: a virus that infects bacteria; also called a phage.
5. Biocatalyst: An enzyme that activates or speeds up a chemical reaction.
6. Biocatalyst: is a container used for bioprocessing.
7. Bio Processing: A technique in which microorganisms, living cells or their components are used to produce a desired end product. Bio processing of biotech products consists of 2 major processing steps:
1. Up streaming process: refers to the culturing of micro erg an isms to create a bulk bio product. This processing is typically done using cell culture or fermentation.
2. Down streaming process: refers to reparation and processing of bulk bio product into a form suitable for its end-use. Typically, this step involves separation, purification and sterilisation.

8. Callus: A duster of undifferentiated plant cells that have the capacity to regenerate a whole plant in some species. This term is used in tissue culture technique.
9. Clone: a cell or collection of cells containing identical genetic material. Clones are produced from a single parent cell.
10. DNA typing: more popularly called as DNA fingerprinting.
11. DNA probe: A molecule that has been labelled with a radioactive isotope, dye or enzyme and is used to locate a particular portion of a DNA molecule.
12. DNA sequence: .The order of nucleotide bases in the DNA molecule.
13. In vitro: performed in a test tube or other laboratory apparatus.
14. In vivo: in the living organism.
15. Interferon: A protein produced naturally by the cells of our body. It increases the resistance of surrounding cells to attacks by viruses.
16. Interleukin: A protein produced naturally by our bodies to stimulate our immune systems.
17. Gene Amplification: The increase, within a cell, of the number of copies of a given gene.
18. Genome: The total hereditary.-material of a cell.
19. Gene Mapping: determining the relative locations of genes on a chromosome.
20. Plasmid: A small circular piece of DNA found outside the chromosome in bacteria. Plasmids are the principal tools for inserting new genetic information into micro organisms or plants.
21. Vector: The agent used to carry new DNA into a cell, viruses or plasmids are often used as vectors.
22. Transposon: A mobile genetic element that can move from one location in the gene and reinsert at another site.
23. Restriction Enzymes: Bacterial enzymes that cleave DNA at very specific location,
24. Protein Engineering: A technique used on production of proteins with new or artificial amino acid sequences.
25. Polymerase Chain Reaction (PCR): A technique for quickly making many copies of a specific segment of DNA.
26. Anti-Sense Technology: The use of an RNA molecule to block gene expression by interfering with protein production. This technique is used commercially in tomatoes to slow ripening for better shipping and longer shelf life.
27. Gene Expression: The physical manifestation of the information contained in a gene,
28. Pleuripotent Stem Cells: are the multi-potential cell populations capable of developing into various specialized cells and tissues of body such as muscle cells, nerve cells, fiver cells blood cells etc.
29. Stem Cell Lines/Colonies: A stem cell line is a self-replenishing colony of embryonic cells.

Applications of Biotechnology

1. Medicine (Health Care)
• Prevention • Vaccine
• Gene therapy
• Genetic counselling
• Diagnosis • Diagnostic kits and
pathological kits
• DNA probes
• Monoclonal antibodies
• Therapeutics • Antibiotics
• Hormones
• Interferon
• Dotting factor
• Usokinase
• Transcription factor based drugs
• Digonucleotide antisense drugs
• Drug delivery systems.
• Other
• DNA fingerprinting and Applications auto antibody finger-
printing
• Fertility control (oral pills)
2. Agriculture
• Productivity (1) Photosynthesis
improver
(2) Transgenic plants
(3) Tissue culture
(4) Bio fertilisers
• Diversity • Hybrid seeds
• Synthetic/Artificial seeds
• Resistance • Disease, drought, pest
resistant varieties
• Allied Areas (1) Seribiotechnology
(2) Aquaculture
(3) Animal husbandry
(4) Food biotechnology
3. Environment and Energy
• Bioremediation
• Bio indicators
• Biodegradation
• Biomass production
• Bio fuels
4. Industry
• Fermentation products
• Mining (Bioleaching)
• Vaccines
• Steroids
• Chemicals
• Vitamins
5. Biotech Instruments
• Biosensors
• Cell analyser
• DNA/peptide synthesisers

Ques. 16 : Briefly discuss the main thrust of activities of India in crop biotechnology?

Ans. Crop Biotechnology : Main Thrust of activities: Agriculture is the mainstay of the Indian Economy. Agriculture and Allied sectors contribute nearly 25 per cent of Gross Domestic Production (GDP), while about 65-70 per cent of the population is dependent on agriculture for their livelihood. India has achieved a major breakthrough in agricultural production as a result of technology evolved by Indian scientists and its wide adoption by the farmers. However, India would need to increase the food production to 250 millions tonnes by the 2020. Therefore, there is a need for critical appraisal of the post-green revolution technology to draw up a balance sheet so that strategies are evolved for achieving sustainable agriculture. The intensive agriculture in India has caused widespread changes in the agro-ecosystem. The insect pest, disease and weed complex has undergone a tremendous change. There is an overall depletion in soil fertility as a result of intensive cultivation of crops. Poor productivity levels and inadequate investments in infrastructure in the last two decades were masked by the huge surpluses, caused partly by low food grain buying capacity and change in lifestyle patterns shifting food patterns. Presently Indian agriculture appears to have reached a plateau and policymakers have been emphasizing the need for new initiatives to boost agricultural outputs. This would not only be essential for ensuring healthy economic growth in the country but also for improving the livelihoods of the poor in rural India.

DBT's Ongoing programme: The department is supporting three main types of research activities under the scheme; R&D projects in priority crops, multi-institutional projects and Plant Molecular programmes in certain institutions/universities. The multi- institutional project involves interaction amongst a number of institutions with well defined objectives. The project on (a) the development of transgenics crops - cotton, rice mungbean and tomato resistance to biotic stresses (b) the project-on molecular characterization and field trials of mustard transgenics for hybrid seed production and low - till cultivation is being pursued by three institutions (c) development of varieties with durable resistance to leaf and stripe rust using molecular marker technology in wheat. This project deals with validation of genetic stocks for documented genes conferring seed and adult plant resistant to leaf rust and stripe rust, identification of molecular markers for important gene and pyramiding important APR and seed resistance genes in common cultivars and (d) Salinity and dehydration stress tolerance in rice: cloning of responsive genes, their promoters and development of transgenics. In this project vectors with proper promoters and genes of interest will be constructed for rice transformation. Finally to utilize the known genes isolated by the participating groups and develop protocols for transformation of rice cultivars. The Plant Molecular Biology programmes are ongoing at MKU, Madurai; TNAU, Coimbatore; Bose institute, Kolkata and UDSC, New Delhi,
Indian initiative for rice genome sequencing: Under Indian initiative for Rice genome sequencing, India has successfully decoded the genome information of the rice chromosome number

11. The rice genome has been shown to have 37,544 genes, of which 1443 genes have been identified in the region sequenced by Indian Scientists. Three projects in the area of Crop biofortification have been sanctioned. Biofortification of wheat for micro nutrients through conventional and molecular breeding approaches, Rice biofortification with enhanced iron and zinc in high yielding non- basmati cultivars through marker assisted breeding and transgenic approaches and development of micronutrient enriched maize through molecular breeding. Under Indo- US collaboration in agriculture biotechnology, two network projects have been recently sanctioned on "Development and evaluation of salt and drought tolerant transgenic rice". DRR Hyderabad, GSSRI, Kamal, ICGEB, New Delhi and Cornell University, Ithaca are partners in the project. The other project is “Fruit and Shoot Borer Resistant Eggplant” implemented at IIVR,Varanasi; UAS, Dharwad and TNAU, Coimbatore

Ques. 17 : Discuss in brief the main thrust of activities and achievements in environmental biotechnology?

Ans. Environmental Biotechnology : Main thrust of activities Realising-the tremendous potential of biotechnology to offer unique, efficient, ecofriendly and economically viable options for waste treatment in situ and degradation of hazardous toxic waste into relatively less harmful or harmless byproducts, the Department of Biotechnology has given a major thrust to programmes for ecorestoration of degraded ecosystems, mining spoil dumps, development of biosensors for detection of pollutants, treatment of industrial effluents, use of molecular markers for characterisation of biodiversity.

The major achievements are:

(a) Establishment of a Laboratory for conservation of endangered animal species CCMB, Hyderabad The Department has established a Laboratory for conservation of endangered animal species at GGMS, Hyderabad jointly with MOEF and AP State Government.

(b) Network programmes on pesticide degradation Three network programmes on degradation of chloro and nitro pesticides in contaminated soils and stocks of banned pesticides involving 4 research institutions each have been supported.

(c) Programme on biodiversity conservation of North Eastern Region Six projects for ex situ conservation, micro propagation and in vitro conservation of rare and endangered plants of medicinal importance, orchids, ethno botanical plants and microbial diversity of north eastern region have been supported.

(d) Programmes for conservation and use of lower plants as indicators of pollution Five projects on biosystematics and conservation studies of liverworts, genetic diversity of ferns, lichens and their use as indicators of pollution have been supported.

(e) Programmes on molecular biology for environmental amelioration. Six projects on characterisation and molecular analysis of polyaromatic hydrocarbon degrading Pathways, genetic engineering for improved heavy metal tolerance, cloning and characterisation of metal resistant genes have been supported.

(f) Ecorestotation of mine dumps and other degraded ecosystems. An integrated biotechnological approach (ISA) for bioremediation of mine spoil dumps and degraded ecosystems has been developed by scientists at National. Environmental Engineering Research Institute (NEERI), Nagpur and University of Delhi and has been successfully demonstrated at a number of sites.

(g) Mangrove afforestation through application of classical and biotechnological tools MSSRF, Chennai has employed an integrated approach combining classical and biotechnological tools to restore and rehabilitate mangrove forests and biodiversity in coastal areas by multi-species enrichment planting. The plantations comprised genetically superior candidate plus-trees identified by various morphological and physiological markers, and these were propagated through a sexual methods (vegetative and tissue culture propagation).

Ques. 18 : Briefly discuss the main thrust of activities and achivements in food biotechnology?

Ans. Food Biotechnology : Main thrust of activities: The Task Force on Food Biotechnology considered R&D proposals in various areas of food processing using biotechnological approaches. These included preparation of tropical fruit juices: food additives (flavours, colourants etc.); large scale production of various species of mushrooms; food from seaweeds; Xanthan gum and on areas of food safety. Development of low cost nutritious food supplements for malnourished school going children was also taken up. Besides, steps were taken to develop diagnostic kits for rapid identification of food borne pathogens. A network programme on the modification of oils and fats using biotechnological approaches was initiated.
Achievements/Leads Obtained/Technologies Developed: The Biotechnology research for food safety resulted in leads for the development of detection kits for various microbial toxins, contaminants and toxicants, PCR assays have been perfected for rapid detection of various food borne bacterial, viral and parasitic pathogens.
Technologies transferred/commercialized: The technologies ‘already commercialized' are as follows: Technology for large-scale production of oyster mushrooms was developed, for processing of 300 kg fresh mushrooms per day. Technology for Spirulina production with an estimated production capacity of 50 kg per day was developed

Ques. 19 : Briefly discuss the major achievements in Human Genetics and Genome analysis?

Ans.

1. Main thrust of activities:
The programme was initiated with the main objective to develop basic capabilities, strengthen existing Institutions which have good expertise, to initiate work in molecular genetics and to support some application oriented projects so as to reduce the burden of genetic disorders in the country. The proqramrne also aims at identifying, mapping and characterising genes associated with genetic disorders and diseases prevalent in India and exploit in the knowledge created by the available human genome, microbial and parasitic genomic sequences so as to usher in molecular medicine tools for better management of disorders and diseases.

2. Achievements : Since its inception of this programme, the Department has implemented several projects; including genetic diagnosis cum counseling units to provide diagnosis and counseling to the affected families for the common genetic disorders prevalent in the country. Major programmes have been initiated in the area of human genetics, functional genomics, human genome diversity, gene therapy, microbial and computational genomics, structural genomics, biocomputing, DNA micro array facilities, clinical proteomics, pharmacogenomics etc, involving large number of clinicians, molecular geneticists and anthropologists. A strategy plan/roadmap document for the 11th P1an was prepared to initiate major programmes in human genetics and genome analysis including genetic education in the country.
Functional Genomics: Under the "Programme on Functional Genomics" implemented at Institute of Genomics & Integrative Biology (formerly CBT), Delhi established good linkages with clinicians in the three major areas i.e. spinocerebellar ataxia (SCA). schizophrenia and biploar disorder and asthma. Developed high throughput capabilities and highly skilled manpower in the field of genomics and computational biology. The considerable progress has been made to carry out genetic research in SCA, schizophrenia & bipolar disorder and asthma. Susceptibility on locus on chromosome 22 for schizophrenia and biopolar disorder in the Indian population has been identified. A US patent was filed for a method to detect human spinocerebellar ataxia 2 gene variants.
Human Genome Diversity: India is one of the first country to take up the project on "Human Genome Diversity". A major consortium project has been implemented at ISI, IICB and SINP, Kolkata. DNA typing of 25 autosomal markers in 53 ethnic groups (tribal, casts and religious communities) of India has been completed. This study also provided evidence of human migration from India to South-East Asia. It revealed that the expansion of the ethnic population took place about 50,000 years ago. The studies indicate that austro-asiastic speaking tribal population in India were the original inhabitants. The group also generated mitochondrial DNA (mtDNA) polymorphism through restriction fragment length polymorphisms (RFLPs) and sequence data on several ethnic populations of India. Utilizing the DNA sequence data of the hypervariable segment 1 of the mtDNA, the group has demonstrated that the tribal population of India underwent a major demographic expansion in prehistoric times.

Ques. 20 : Briefly discuss the main thrust of activities in Medicinal biotechnology?

Ans. Medicinal Biotechnology : Main thrust of activities: Medical biotechnology and health care areas have emphasis on: molecular diagnostics, genetic vaccines, molecular characterisation of pathogens, new targets: and novel drug delivery systems relating to viral, bacterial and parasitic infections markers for cervical and oral cancers, stem cell biology, development of products and processes, and development of advanced infrastructure and expertise.
The priorities of the programme for health and health related biotechnologies have been taking into consideration the criteria like: Disease burden; Cost effectiveness of technologies. Considering emerging and reemerging diseases; Disorders because of prevalent life style and potential utilities of these technologies at community level. Research on reproductive human health and contraception with special-interest for developing along-term strategy for immunological control of reproduction both in males and females. Edible vaccines for infections like cholera, hepatitis, rabies contraception etc.

Ques. 21 : Give an account of the achievements in vaccines?

Ans. Vaccines
A. IMMUVAC
Achievements/leads/Technology: An immuno-modulator Based on killed Mycobacterium developed by the National Institute of Immunology, New Delhi to be used as an adjunct to MDT and for prophylaxis in leprosy patients. Upscaled and commercialized by M/s Cadila Pharmaceuticals, Ahmedabad, initially as LEPROVACr, now as IMMUVACr It received Orphan drug designation in USA as an adjuvant to MDT for multi-bacillary leprosy. Recently it has shown a cure rate of 98% in category-II Tuberculosis patients in contrast to a cure rate of 56% with MDT under DOTS (as per WHO report).
B. Rabies
Main thrust of activities: (i) Project Title: National Jai Vigyan Mission Programme on Rabies; (ii) Objectives: Preclinical toxicity and protective efficacy of a Rabies DNA vaccine in Mice, Monkeys and Dogs – IISc, Bangalore; Indian Immunologicals Ltd. & NIN, Hyderabad.

Ques. 22 : Briefly discuss the achivements in the field of plant biotechnology?

Ans. Plant Biotechnology : Plant Biotechnology has been one of the thrust areas of the Department and major programmes have been supported on Forest tree, horticulture and plantation crops. The thrust is on application of tissue culture for regeneration of high quality economically important plant species, demonstration of large-scale plantation and validation of proven technology; germplasm characterization, improvement of crops through molecular biology tools, basic research, genomics initiative; host pathogen interaction, resolving of taxonomic problems by molecular interventions etc.
Achievements vis-a-vis goals/targets.
R&D: Studies were carried out for developing/standardization of micro propagation protocols of important tree species. Under the horticulture crops, post harvest studies for improved shelf life of Tomato, Grapes and Banana were supported. Transformation systems have been established. Transgenics of tomato are undergoing field trial. Improvement of selected spices (black pepper, cardamom, ginger, vanilla) through biotechno-logical tools were supported concentrating on germplasm characterization and conservation and screening for disease resistance, developing improved varieties through transformation and mutation breeding.
Solanaceae Genome Initiative: An International Solanaceae Genome Initiative has been launched with the aim of creating a coordinated network of knowledge regarding the Solanaceae family and addressed various key issues. The long-term goal of the SOL programme is to create a network of map based resources and information and address key questions in plant adaptation and diversification through this International effort. India Joined the International 'SOL' programme and has sequenced chromosome 5 (12 Mb).
Basic Research: The thrust of the programmes supported under Basic research was to study the signal transduction pathway in identified plants to understand the mechanism involved in responding to extend stimuli and the process of adaptation to these changed environment conditions. The study aims at identifying the components of Signaling pathway with respect to (a) early signal transduction (b) transcription factors involved in gene expression and (c) molecular events leading to protein turn aver. The role of hormones light and stress studies on plant behaviour, floral development and root differentiation.
Impact: Tissue Culture Protocols developed for Important crops - Apple, Citrus have been developed for the first time. National Certification System for Tissue Culture plants has been developed for the first time.

Ques. 23 : Discuss in brief the major scientific achivements in seri biotechnology?

Ans. Seri Biotechnology : The Department of Biotechnology (DBT) is implementing a programme on application of biotechnology for increasing productivity, enhancing silk quality and improvement of host plants in both mulberry and non-mulberry sericulture.
Major Scientific Achievements: Work accomplished on identification and characteri-zation of silkworm microsatellites and utilization of microsatellite derived markers for strain typing, phylogenetics, linkage mapping, Z-chramosome mapping and analysis of heterologous silkmoths. Application of these markers for identification of productive silkworm hybrids in combination with conventional breeding carried out. India joined "International Consortium on Lepidopteron Genomics" and fulfilled the Indian commitments viz (a) Construction of high-density linkage map of silkworm (Bombyx mori) (b) Isolation and characterization of ESTs from non-mulberry silkworms. Microsatellite markers for tasar silkworm and muga silkworms have been developed which are being, used to study population structure, gene flow and genetic polymorphisms. Characterization of anti-bacterial proteins in silkworm has led to the discovery of unique insect specific lysozyme (similar to C-type lysozyme and alpha lactalbumins). A Web-enabled DNA database on mulberry has been developed for the first time in the world encompassing DNA fingerprinting studies carried out on more than 250.genotypes including 50 selected- elite genotypes. Efforts have been initiated for construction of a framework molecular linkage map of mulberry for mapping and identification of useful genes. A Multi-institutional Network project has been launched on identification of DNA markers associated with disease and pest resistance in mulberry involving CCMB, Hyderabad alongwith four institutions of CSB and state governments.

Ques. 24 : What were the goals for seri biotechnology in the eleventh plan?

Ans.
Goals for 11th Plan:
• To develop novel high yielding silkworm varieties producing good quality silk and resistant to diseases through marker-assisted breeding and transgenic route.
• To develop biotic / abiotic stress tolerant mulberry varieties through marker assisted breeding and transgenic route.
• To support more basic research in mulberry sericulture (both silkworm and host- plants) including research on disease surveillance and casual agents of major disease of non-mulberry silkworm.
• To develop suitable technology(s) for by-product utilization in sericulture industry and value addition to the system toward better economic gains. To develop various applications of silk proteins (both sericin and fibroin) for medical and cosmetic purposes besides using pupa as a source of protein and bio-fuel.
• To develop silkworm as bioreactor for producing high-value proteins.
• To promote HRD in the area of seribiotecnology by following novel and integrated approaches.

Ques. 25 : Briefly discuss the main thrust of activities and achivements in stem cell research?

Ans. Stem CelL Research : Main Thrust-of Activities: The understanding of stem cells biology has opened up new medical paradigm especially in tissue generation as embryonic stem cells could be most powerful ones of all. Pluripotent stem cells are the multi-potential cell - populations capable of developing into specialized cells and tissues of the body such as muscle cells, nerve cells, liver cells and blood cells and may be used for replacement of cells and tissues to treat many diseases and disorders like Parkinson's disease, spinal cord injury, stroke, burns, heart disease, diabetes, osteoarthritis, and rheumatoid arthritis and also help us understand what causes birth defects and cancer. The development of stem cell lines that may produce many tissues of human body is an important scientific breakthrough. Recent studies, however, have clearly led to development of technologies where by the stem cell either from the adult or from the embryo can be isolated and grown in the tissue culture. This discovery has surprised many scientists and this technology is now being utilized by several scientists specially in the developed countries for organogenesis, transplantation of the stem cells to, rectify the ailing and disease tissue, delivery of new therapeutics, delivery of interleukins to modulate immune responses and also delivering candidate vaccine for several infectious diseases. This research has the potential to revolutionise the practice of medicine and improve the quality and the length of life.
Achievements: A number of projects have been implemented on use of limbal stem cells in repairing damaged ocular surface; neural stem cells to understand how stem cells get. differentiated into different types of neural cells, haematopoietic stem cells, isolation, characteri-zation and neuronal differentiation of human embryonic stem cells. In addition, the department has also received the programmes on other aspects of stem cell biology. Significant achievements are: Many patients have undergone cornea transplantation using limbal stem cells. First haploidentical stem cell transplantation has been done. For this, CD34+ cells have been collected from the father of a child suffering from glanzmann thrombasthenia who had severe persistent bleeding. This type of transplantation is first of its kind in India.
Proposed Programmes: To establish stem cell banking facility in the country. To develop well characterized stem cell lines. Potential use of embryonic stem cell lines to generate various organs. Regeneration of tissues for stem cells obtained from aborted foetus. Basic understanding of the signaling mechanisms and differentiation of stem cells.

Ques. 26 : What is cloning?

Ans. Cloning is the production of identical animals, plants or microorganisms from a single individual. A clone is an organism that is derived from a single parent through non-sexual activities.
Nature itself is the greatest cloning agent. In one out of 45 human conceptions the fertilized ovum splits for some unknown reason and produces monozygotic (identical) twins. Each has a genetic make up identical to the other. In cloning this operation is done intentionally in a laboratory.
It must be noted that natural cloning is common on plants, microorganisms and simple animals such as corals. Many organisms which reproduce a sexually produce their own clones. But mammals which reproduce sexually cannot clone naturally. The progeny of a mammals inherits genetic matter not from one parent but half each from both parents. therefore the younger produced is never an identical copy of anyone of its parents. Natural clones in mammals are confined to the production of identical clones.

Answer of Question 1: B

Answer of Question 2: B