Biotechnology -Implementation of Biotechnology

The Biotech On web is a blog for those student who are interested to make their career in food technology or biotechnology.Food technology is the study about food processing and their key terms,In food technology we know about various branches of food study.Today biotech is one of the most successful choice of science student. Many Universities offered course named "Food Chemistry".In my opinion food technology is okay for teaching purpose, but today many multinational company which deals in food product that offered job of successfully qualified in graduation (food chemistry) with handsome package.

This is recent branch of biology. Biology in relation to technology is called biotechnology or biotechnology is defined as application of living organisms or their processes in manufacturing industries.

Study of use of living and the substances produced due to their activites is called biotechnology.
Mostly micro-organisms are used in many industries as alcohol, enzymes, vaccines, vitamins, antibiotics, organic acids, etc. So, biotechnology is controlled use of microbes for benefits of human being. In this science, principles of biochemistry, molecular biology and microbiology are mostly  used. Some important examples of biotechnology products are:

1. Alcohol: It is result of yeast fermentations, which is the incomplete oxidation of complex organic compounds with the help of enzyme produced by yeast (invertase and Zymase)

In the middle of the 19th century, Louis Pasteur reported that alcohol,beer and butter milk are the result of yeast fermentations .Yeast fermentations is the basis of baking as well as brewing industries, besides preparations of fermented foods like idli, dosa, etc. Saccharomyces cerevisiae is commonly called baker's yeast or brewer's, is a saprophytic unicellular ascomycetes fungus growing on sugary solution.  

Some other common products of yeast fermentations are:
Beer - It is produced from Hordeum vulgare (barley) malt and alcohol content is 4-8%.

Wine: Produced from grapes by fermentations and alcohol content is 10%- 20 %

Brandy :- Produced by distillation of wine and alcohol content is 43% - 57%.

Gin: Produced from fermented European rye, i.e, Secale cereale.

Rum : Produced from molasses of sugarcane and alcohol content is 40%.

Thus the type of the alcohol depends upon medium and agent causing fermentations. The species involved in above case are: Saccharomyces cerevisiae, S. ellipsoides, etc.

2. Organic acids: Some organic acids are produced by fermentation caused by fungi and bacteria.Some important are:

a. Lactic acids: Produced by fermentation of corn starch, molasses, potato, whey, etc, by Lactobacillus bulgaricus, L. delbrueckii and streptococcus lactis.

b. Acetic acids(Vinegar ): Produced by two step fermentation of sugarcane juice by yeast and Acetobacter aceti bacteria.

c. Citric acid: Produced by aerobic fermentations of sucrose in beet molasses by Aspergillus niger fungus.

d. Gluconic acid: Produced during oxidation of glucose by fungi like Aspergillus, penicillium, Mucor, etc.

e. Fumaric acid : Produced from sugar by activity of Rhizopus nigricans (bread mould)

3. Enzymes: The term 'enzyme' was first of all used by William Kuhne (meaning----in yeast). Enzymes are the organic substances which enhance the rate of biochemical reactions (without themselves undergoing any change). Enzyme can also be used outside the cell  to catalyse some specific biochemical reactions to manufacture many industrial products. Out of total over 2200 enzymes known today,only a few are used in industries, medicines and food which are as follows.

a. Proteases or proteolytic enzymes: obtained from Aspergillus oryzae and Bacillus subtilis and are used in detergents to remove some proteinaceous stains  on clothes.

b. Amylases: Obtained from Bacillus subtilis. Aspergillus niger.A oryzae, etc. and are used in beer and bread making. Also used for softening starched clothes.

c. TPA (Tissue plasminogen activator): This enzyme is specifically used in heart patients to dissolve blood clots.

d.papain: This protein digesting enzyme obtained latex of papaya,is used for making meat tender.

e. Lactase: Obtained Saccharomyces fragilis and Torula cremori andused for making cheese from pasteurized milk and also for preventing lactose crystals in ice-cream.

f. Invertase: Obtained from Saccharomyces cerevisae  and is used to prevent sandliness in diary products.

g. Thrombin: obtained from beef plasma and used for blood clotting in surgical operations.

h. Renin (Rennet) : Danish chemist Christian Hensen (1874) first obtained it from calf stomach and used for cheese formation

i. Pectinases: Obtained from Asper gillus lucherisis and used for destruction of penicillin action.

j. Cellulase: Obtained from Myrothecium verrucaria and used in production of dextrin and fructose.

Enzymes are more fragile than inorganic catalysts.Further these are more expensive and are rapidly deactivated at high temperatures.Stabilisation of enjymesis done by enzyme immobilization,which is done by fixing it to solid support,entrapping in a gel,cross-linking the enzyme molecules or by encapsulation of enzymes in small artificial cells.This immobilisation protects enzymes against attack by proteases.Further immobilizsed enzymes can be easily separated from reaction mixture at the end of reaction and then can be reused.

4.Vitamines:Vitamines are the organic compounds,provided to the body with diet and are required for some biochemical processes in very small amounts.Vitamines were discovered by C.FUNK(1911) and different types of diseases
(like beri-2,scurvy,rickets,night blindness,etc)are produced as a result of vitamines deficiency.Some microbes are good source of vitamines and the first microbiologically produced vitamin are vitamin c (ascorbic acid) during this fermentaion with a wild strain of bacteria (by Gyorgy).After this different microbes are being exploited for commercial production of different vitamines.Some important examples of microbes-produced vitamines are:

a.Riboflavin:(vit.B2):It is synthesised for industrial purposes by fermentation with a filamentous yeast, Ashbya, gossypii. The main sources of vit. B2 are cereals, yeast, leafy vegetables,milk,etc. It is precursor of FMN and FAD, which act as coenzymes for dehydrogenases enzymes.

b.Cobalamine (vit. B2): This vitamin is of animal protein origin like liver, meat, fish. It is not present in vegetarian diet and its deficiency causes pernicious anaemia. Now a days this vitamin is being produced by fermentation using Pseudomonas denitrificans, Bacillus coagulans, B.megatherium and propionic acid bacteria, i.e., Propionibacteria.

c. Precursor of vitamin A, i.e carotene is produced by fermentation using Blakslea trispora.

d. Precursor of vitamin C, i.e. L - sorbose by Glucanobacter oxidans.

5. Antibiotics: These are the substances of microbial origin and having antimicrobial activity or these are the chemical substances secreted by one micro-organisms, which inhibit or check the growth of other micro organisms.First antibiotic discovered was penicillin(wonder drug), from penicillium notatum by sir Alexander Fleming (1928) However, the term 'antibiotic' was given by waksman  (1942). Waksman himself discovered two antobiotics i.e, Actinomycin (1941) and Streptomycin (1942) . Main sources of antibiotics are fungi,
bacteria and actinomycetes and about 7,000 antibiotics are known at this time. There are two main groups of antibiotics.

a. Broad spectrum antibiotics are those which inhibit the growth of many pathogenic species, differing from one another in their structure.

b.Narrow spectrum antibiotics are those antibiotics which inhibit the growth of a specific pathagenic species.
Different antibiotics have different modes of action, e.g., by destruction of cell membrane, inhibition of cell wall synthesis or checking protein synthesis or inhibition of nucleic acid synthesis.

Common antibiotics are penicillin . streptomycin, Terramycin, Aureomycin, Chloromycetin or Chloramphinicol, Griseofulvin, Neomycin, Erythromycin, Polymyxin, Subtelin, etc.

Streptomyces griseus strains are known to be producers of 32 different structural types of bioactive compounds (antibiotics)

Antibiotic of algal origin is Chlorellin and of lichen origin is Usnic acid.

6. Yoghurt: It is a special diet, which is prepared by fermentations of concentrated milk first with Lactobacillus bulgaricus and streptococcus thermophillus at 40-46 degree C for 4 hrs. and then fermentations with yeast.

7.Gibberellins: This plant growth hormone or phytohormone is obtained from a fungus called Fusarium moniliformae (or Giberella fujikuroi). This was isolated by Yabuta and Sumuki (1939). At this time, over 52 gibberellins have been discovered  from different plants.

8.Steriods: Steriods are molecular weight fatty compounds, which are having one 5-carbon ring and three 6-carbon rings. Steriods are of wide distribution in both plants and animals . Chlolesterol is one of the most important steriods of animals, which is a precursor of animal hormones (steriods hormones) like progesterone and estrogen (female sex hormones) and testosterone (male sex hormone).

Progesterone is precursor of 4 different useful steriods and the micro-organisms used in this conversion are Rhizopus arrhizus, R. Stolonifer (R. nigricans), Curvularia lunata (all fungi) and a bacterium streptomyces argenteolus.

9. Insulin: Insulin is  a hormone of protein nature (51 amino acids),made of 2 polypeptide chains having 21 and 30 amino acids respectively and joined by disulphide nonds. This hormone is produced be cells of 'Islands of Langerhans' of pancreas and is responsible for controlling blood sugar level. Deficiency of this insulin leads to a disease called diabetes mellitus. Insulin supplied from outside can cure this disease.
Banting and Best(1921) first of all isolated insulin from dog's pancreas and used it for curing diabetes patients, Insulin is now obtained from pancreas of slaughtered pigs and cattle.This insulin slightly differs from man's insulin and effectively controls diabetes.
By using genetic engineering or recombinant DNA technology, insulin producing genes from human beings have been transferred into E. coli bacteria, which produce insulin called 'Humuline' for clinical use. This type of synthetic insulin was produced by an American pharmaceutical frim Eli-Lily on july 5, 1983.

10. Interferons: Interferons (antiviral proteins) were produced by Charles Weismann of Zurich University through recombinant DNA technology in E.coli in 1980.

11.Monoclonal antibodies: Antibodies are the specialized proteins, which are produced inside the host body in response to foreign substances called antigens (mostly protein or polysaccharides) and provides immunity to the host against antigens. Antibodies are produced in Lymph nodes, spleen and liver.

Monoclonal antibodies (Mabs) are the specialized antibodies, which are specific to only one type of antigens. The concept of monoclonal antobodies was put forward by georges Kohler and Cesal Milstein in 1974 and these antibodies are produced outside the body by hybrid cell culture technique, known as Hybridoma technology. For this technology, Kohler and Milstein were awarded Nobel Prize in 1984.

In this technique, the antigens (against which antibodies are needed ) are injected into rat is removed and from spleen, some lymphocytes (which are responsible for antibody production) are isolated. These lymphocytes are mixed  with myeloma cells (tumour cells isolated from cancer of bone marrow). Some of these lymphocytes  and myeloma cells fuse together to form hybridoma cells (by somatogamous fusion). These hybridoma cells have capacity of antibody production  (of lymphocytes) and rapid cell division (of cancer cells). These hybridoma cells in cultural conditions produce large quantities of specific and pure (monoclonal) antobodies, which are separated and used in cure of different diseases.


12. Vaccines: Production of antibodies against antigens, is the basis of immunity. Vaccine in fact is dilute dose (suspension) of antigens, which is used for developing artificial or acquired immunity . The process of inoculation of vaccine is known as vaccination  and Louis pasteur (1850) is responsible for understanding the basis of vaccination  and immunization. (Edward Jenner 1970, however made earlier studies in relation to small pox). A vaccine contains either weakened (polio, smallpox and measles vaccines)or even killed pathogens( typhoid vaccine ) which have still antigens to induce antibody production. In some cases (like botulism and tetanus), toxins produced by pathogens serve as vaccines . A lare number of vaccines (called first generation vaccines) against different viral and bacterial diseases have been produced.

In recent years, some new vaccines called second generations and third generations vaccines have been developed . Second generations vaccines are those, which are produced by recombinant DNA technology or genetic engineering . e.g., vaccine for herpes virus and Hepatitis B. Third generations vaccines are produced synthetically or are synthesized vaccines, e.g., for feline leukemia virus and foot  and mouth diseases  virus.

13. Amino acids : An important basic amino acid, lysine is produced from diaminopimelic acid. (constituent of cell wall of bacteria, e.g., E.coli by activity of bacterium Enterobacter


15. Some other food products : Cheese is produced by activity of different species of lactobacillus and Streptococcus.
Penicillium camembertii and P. roquefortii are used to give flavour to cheese (camembert cheese and roquefort cheese).
Similiarly, butter is produced from cream by activity of streptococcus lactis and Leuconostoc species.

15.Dextran : Sucrose by activity of Leuconostoc mesenteroides bacteria, gives rise to a complex polysaccharide called dextran, which is used in plasma tranfusions.

16. Cholesterol lowering statins: Monascus purpureus (red yeast ) is a species of mould that is purplish red in colour and is known by the names ang-khak rice mould, corn silage mould, maize silage mould, and rice kernel discoloration. This fungus is most  important because of its use in the form of red yeast rice and in the production of certain fermented foods in china. However, discovery of production of cholesterol  statins bythe mould has prompted research into its possible medical uses. It produces a number of statins. The naturally occuring lovastatins and analogues are monacolins K, L,J and also occur in their hydroxyl acid forms alongwith dehydroxymonacolin and compactin (mevastatin).The prescription drug lovastatin, identical to monacolin K, is the principal statin produced by Monascus purpureus.

17. Tissue cultue: This is also a latest method of crop improvement. Besides also used in manufacture of antibodies, alkaloids, and dyes (like shikonin, a red dye used in silk industry and in treatment of burns is obtained from culture of cells of lithospermum root)

18. Single cell proteins (SCP): This terms was first used in 1966. But this term is misleading now a days because it denotes not only the isolated cell protein but it is used for any microbial biomass from uni-multi-cellular bacteri, yeast, algae or filamentous fungi which can be used as food or food additives. In general, microbial biomass contains 45-55 % protein, though in some bacteria, upto 80% protein content present, Large scale production of microbial biomass (SCP) has advantages over traditional methods of producing proteins as microbes have high rate of multiplicationm, have high protein content, can utilize large number of different carbon sources (waste products also) and microbial biomass production is independent od seasonal and climatic variation. Most commonly used microbes are Spirulina (blue green alga), Chlorella (green alga), Saccharomycopsis) lipolytica (fungus), mushrooms, etc.

Methlophilus methylotrophus bacteria have been investigated for use in single cell protein production. These bacteria have a generation time of about 2 hours and mainly used in animal feed as bacteria, in general produce a more favourable protein composition than yeast or fungi. Therefore, the large quantities of SCP animal feed using bacteria. The resulting product appears pinkish white, is odourless, tasteless and non-toxic which consists of about 65-75% protein of balanced amino acids composition.

Probiotics are live microorganism thought to be beneficial to the host organism. This term has been derived from latin ('pro -for+ biotics, derived from noun 'Bios-Life). According to currently adopted definitions by FAO/WHO, probiotics are live microorganism which when administered in adequate amounts confer a health benefits onthe host. Lactic Acid Bacteria (LAB) and bifidobacteria are the most common types of microbes used as probiotics, but certain yeasts and bacilli may also be helpful. Probiotics are commonly consumed as part of fermented foods with specially added active live cultures; such as yoghurt, soyyohurt or as dietary supplements. At the start of 20th century, probiotics were thought to beneficially affect the host by improving its intestinal microbial balance, thus inhibiting pathogens and toxins producing bacteria.

19.Sewage treatment and methane production using microbial systems: Conventional sewage treatement involves the use of the naturally developing micro-organisms within the sewage treatement system. Now a days the sewage is onoculated with a specific micro organisms, specially selected for that particular sewage treatement process. These organisms are called ' Starter cultures'

A strain of Pseudomonas putida containing plasmids has been developed, which degrade octane, xylene, metaxylene and camphor.

Methane is produced during anaerobic decomposition of sewage and other organic wastes by bacterias. The methane is collected and used as fuel in many countries.

Fringe benefit of Biotechnology

The application of Biotechnology to unravel the atmosphereal issues within the environment and within the ecosystems is named Environmental Biotechnology. it's applied and it's wont to study the natural atmosphere. consistent with the international Society for atmosphereal Biotechnology the environmental Biotechnology is outlined as AN environment that helps to develop, with efficiency use and regulate the biological systems and forestall the atmosphere from pollution or from contamination of land, air ANd water have work with efficiency to sustain an atmosphere – friendly Society. In alternative words, it may be outlined because the best use of the natural resources through the plants, animals, bacteria, fungi and alga so as to supply the energy that comes from the resources that area unit natural like daylight, wind, air, waves (renewal in a position energy), food and nutrients. The best use of the these natural resources may be done by re-cycling and utilizing the waste from these resources and realize distinctive ways that of making methodes to utilize the waste and wherever the waste of every method created becomes the feedstock for one more process.

Application and advantages of Environmental Biotechnology:

Applications:
There area unit 5 major differing kinds of Applications of Environmental Biotechnology. they're as follows:

a) Biomarker:
This type of Application of environmental Biotechnology offers response to a chemical that helps to live the extent of injury caused or the exposure of the harmful or the pollution impact caused. In alternative word, Biomarker may also be known as because the Biological markers the foremost use of this applications helps to relate the affiliation between the oils and its sources.

b) Bioenergy:
The collective purport of Biogas, biomass, fuels, and H area unit known as the Bioenergy. the employment of this application of atmosphere Biotechnology is within the industrial, domestic and area sectors. As per the recent would like it's finished that {the would like|the necessity|the requirement} of fresh energy out of those fuels and alternative routes of finding clean energy is that the need of the hour. one amongst the pioneer samples of inexperienced energy area unit the wastes collected from the organic and biomass wastes; these wastes facilitate use to over the pollution problems caused within the atmosphere. The Biomass energy provide has become a distinguished importance in each country.

c) Bio-remediation:
The method of improvement up the risky substances into non-toxic compounds is named the Bioremediation process. This method is majorly used for any quite technology pack up that uses the natural microorganisms.

d) Bio transformation:
The changes that occur within the biology of the atmosphere that area unit changes of the complicated compound to straightforward non-toxic to harmful or the opposite means spherical is named the bio transformation method. it's employed in the producing sector wherever harmful substances area unit born-again to Bi-products.

Benefits:
The major advantages of environmental biotechnology area unit it helps to stay the environment safe and clean for the employment of the long run generations. It helps the organisms and also the engineers to search out helpful ways that of obtaining tailored to the changes within the atmosphere and keep the atmosphere clean and inexperienced. The good thing about atmospherically biotechnology helps United States to avoid the employment of risky pollutants and wastes that have an effect on the natural resources and also the environment. the event of the society ought to be exhausted such the way that it helps to safeguard the environment and conjointly helps United States to development it.

The environmental biotechnology includes a role to play within the removal of the pollutants. it's turning into a plus for the scientists and also the environmentalists to search out ways that to convert the waste to re-useable product. The applications of environmental biotechnology are getting a benefiting issue for the environment; the applications includes – genetics, proteomics, bioinformatics, sequencing and imaging processes area unit providing massive amounts of knowledge and new ways that to improvise the atmosphere and shield the atmosphere.

Bio-remediation:
The branch of Biotechnology is named Bioremediation and it deals with the issues associated with the atmosphere. the employment of various forms of contaminants and fungi area unit wont to the clean the atmosphere and it plays a awfully very important role to stay the pollutants off from the atmosphere. The microorganism area unit thought of jointly of the very important microbes since they break the dead organisms or the materials into helpful organic matter and nutrients. As per the analysis not all the contaminants may be touching the atmosphere may be destroyed victimization the method of bio remediation eg. Lead and metallic element aren't the contaminants that may be rotten by the microorganisms.

The process of Bio remediation takes place in two conditions – aerobic and anaerobic conditions. once the microbes would like O to perform its method is within the case of aerobic condition; if they will ample quantity of O they’ll be able to provide most quantity of water and carbon through the conversion of contaminants and toxins. just in case of anaerobic conditions the microbes perform their work while not the presence of O the chemical compounds gift within the soil helps the anaerobic to perform its duties with efficiency.

Types of Bioremediation:

a) Mycoremediation:
This is a kind of Bioremediation; fungi area unit used for the method of remotion. the employment of plant mycelia in bioremediation is named Mycoremediation. The role of the plant within the scheme is to perform the work of braking down the organic substances into abundant smaller and easier materials. The plant structure helps in braking down the substances and that they secrete extracellular enzymes and acids that brakes polymer and cellulose; these area unit building blocks of plant product. The key perform of Mycoremediation is to focus on the correct plant species for a selected waste material.

b) Phytoremediation:
The direct use of the inexperienced plants and their microorganisms wont to balance or decrease the contaminated soils, sludges, sediments, surface water or well water is named Phyto remediation. As per the traditional Greek term phyto means that plant and remedian means that restoring balance. this kind of bioremediation explains the way of treating the environmental issues with the assistance of plants. The part of Phytoremediation consists of contaminated soil, water, and air that area unit contaminated and also the plants area unit able to contain and eliminate the metals, pesticides, solvents, explosives, crude oil.

C) microorganism Remediation:
The use of microorganisms to degrade organic contaminants and to bind the employment of metals in less believable type is named microorganism rectification. – Aerobic and Anaerobic conditions. once the microbes would likeOxygen to perform its method is within the case of aerobic condition; if they will ample quantity of Oxygen they’ll be able to provide most quantity of water and carbon through the conversion of contaminants and toxins. just in case of anaerobic conditions the microbes perform their work while not the presence of Oxygen the chemical compounds gift within the soil helps the anaerobic to perform its duties with efficiency.

Concerning Molecular biotechnology

The study of nucleic acids and proteins exploitation the research lab techniques is named Molecular Biotechnology. It involves a larger scope for varied types of analysis and analysis. the varied types of analysis concerned ar medicine, biology, genetics, cell Biology and biology. The importance of Molecular Biotechnology is changing into an impending method within the field of Agriculture and our surroundings and therefore the importance of Molecular biotechnology is completed by the complete world as per the recent study knowledge. it's the flexibility to transfer genetic data between totally different organisms. the most goal of Molecular Biotechnology is to grasp the various biological processes concerned and creation of wise merchandise. The human ordination project has opened new gates for the biological progress, development within the pharmaceutical business to make innovative medicines to cure diseases and conjointly varied types of treatments and to improvise the present processes concerned in creating medicines within the pharma domain.

Past of Biotechnology

The Study of Macro molecules and molecule mechanisms that has an look within the living organisms is named Molecular Biotechnology. The molecular Biotechnology's main aim is to target the structure and therefore the operate of the factor, nature of the factor, replication of the factor, expressions and mutations of the genes. The construct of Molecular Biotechnology was introduced from the year 1930 ANd 1940 and it had been not that impending at the initial stages of its introduction it solely received an importance within the years Nineteen Fifties and Sixties. the employment of Molecular Biotechnology was done by geneticists, structural chemists and physicists. Francis Henry Compton Crick introduced himself as a biologist and delineated  additional as “a mixture of a crystallographer”.

Key benefits of Molecular Biotechnology:

There ar sure reasons with regards to the advantages of molecular biotechnology; the advantages ar associated with the surroundings. It helps in inspecting the surroundings and it helps in observance the requirements through Molecular biotechnology, it makes the consultants to grasp the environmental, management wants and therefore the risk concerned and evaluates the chance issue with efficiency. Any damages to soil or water it helps to produce treatment. With the assistance of introduction of Molecular applied science and inexperienced Technology the examination is completed additional promptly and with efficiency. just in case of any environmental disputes the method of Molecular Biotechnology plays a crucial role to look at the sort of dispute and helps to produce an acceptable answer to the dispute.

Biotech in Animal agriculture

Biotechnology- Animal Agriculture

There is an enormous impact of Animal Agriculture on the surroundings and therefore the climatically changes. Animal agriculture consumes plenty of our natural resources and helps within the method of deforestation. world organization within the year 2006 came up with a study stating that there's eighteen of contribution globally by the animal agricultural sector. the main drawbacks of animal agriculture square measure that it produces immense amount of animal wastes, damages the water and causes pollution and conjointly pollution. This results in a forceful modification within the climate and therefore the surroundings. per the recent study at each stage of production of raw food like egg, meat and milk merchandise has an effect on the climatically changes and this results in climatically crisis because of the discharge of greenhouse gases within the atmosphere.


There is a serious transformation in biotechnology because of the fast changes and advancement in Animal Agriculture. It involves sure technologies so as to improvise the needs; the assorted styles of technology embody biotechnology, genetic modification, transgenic, varied polymer techniques and therefore the method of biological research. The EFSA (European Food Safety Authority) written a scientific study on the problems with regards to the protection and health of animals, quality of food and therefore the aspects of the surroundings associated with the method of biological research. solely the food created from healthy cloned animals aren't any less safe than the food merchandise created from the bred animals.

Biotechnology- Bio piracy (Bio colonialism)

Biopiracy may be defined as, " The theft of various natural products  (of plants and animals) and then selling them by getting patents or intellectual property rights, without giving any benefits (financial or technological) back to the host country or country of origin."e.g., Biopiracy of Neem, Haldi Basmati rice, etc.

Biopiracy of Neem: Patenting of seed oil from indian Neem (Azadirachta indica fam. Meliaceae) by US Department of agriclulture (USDA) and W.R Grace (Columbia) in 1992, was a case of biopiracy (patent No. 436357 BY). For generations, Indians had used Neem oil as a medicine and pest controlling agent. So the US patent on Neem had created a great controversy. In May, 2000 after a long legal battle, the European patent office (EPO) revoked the patent of USDA and W.R Grace.

Biopiracy of Haldi
In march 1995 University of Missisippi Medical Centre (USA) had taken patent on turmeric powder (curcuma domestica fam. Zingiberacease), as a wound healing agent. But on August 13, 1997, this patent was revoked due to objections of CISR because for a long time, turmeric powder is used as a wound healing agent in india and it was not a discovery of US patent.
Similiar is the case of biopiracy of Basmati rice.

Misuse of intellectual Property Rights (IPR) can lead to biopiracy, e.g,. University of georgia Scientists explore the Oxaca state of Mexico for indigenous plants of medicinal/pharmaceutical potential and patent any plant with promise. The corporations gain, but people who have used it for ages lose it. This is a major threat to the treasure of traditional knowledge in india. South asia.

The problems of ownership and concerns over right of countries were discussed in a documents known as Convention on biological diversity (CBD) in first Earth summit, Rio de janeiro (1992, brazil). CBD provides for sovereign rights for individuals countries on their genetic resources. USA and EU strongly opposed the view held by developing countries that the sovereign rights of the countries of origin of PGRs (Plant Genetic Resources) held in ex situ collections be recognized. These developed countries have been successful in achieving approval for a call to strengthen co-operation to sustain ex situ collections and recognizing that states have sovereign rights over their own PGRs (PGRs stored on their territory irrespective of the territory of origin ) Thus, recognition  of sovereign rights of the countries of origin of PGRs available in gene banks in the developed countries has been rejected by USA ans EU. Therefore the companies present in Northern countries are dealing directly with agencies where PGRs are located rather than dealing with countries  of origin of those PGRs A pharmaceutical industry can purchase tropical PGRs from botanical gardens located in temperate countries. According to international  Association of botanical, 50% of world's vascular plants are available in botanical gardens and 75% of these gardens are present in developed countries on the North. The pharmaceutical industry is exploring the possibility of getting rights for chemical analysis of collections at these gardens for development of new drugs. e.g., Pfizer pharmaceutical Co. (USA) has collaboration with New York botanical gardens and through it has connections with botanical gardens in Hawaii. This is also a case of bio piracy through gene banks and botanical gardens.

Biotechnology - Bio safety issues

Bio patent
With emergence of modern biotechnology, one important issue raised which is the legal characterization and treatment of trade related biotechnological processes and products, commonly called intellectual property . In simple words, intellectual property is a 'product of mind'.
In biotechnology,one of the most important examples of intellectual property is the processes and products, which result from development of genetic engineering techniques through use of restriction enzymes to create recombinant DNA. The rights to protect this property are called intellectual property rights (IPR). Intellectual property rights include rights relating to literary, artistic, scientific works, inventions in all fields, industrial designs, trademarks, commercial names, etc. Intellectual property rights include patents, Copyright, trade marks and trade secrets.

A patents is a government granted and secured legal right to prevent other from making, using or selling the inventions covered by patent. A patent is a personal property which can be licensed or sold like any other property.

Under USA law, a patent means grant of right to exclude other from making, using or selling an inventions for a 17years period. The Indian Patent Act(1970) allows process patents but not product patents. The duration of the patent in india is five years from date of grant of patent or seven years from date of filling the application, whichever is less.

Patenable subject matter should meet 3 requirements as utility, novelty and statutory subject matter. Difference countries have different patent laws and these can be modified from time to time.Complaints of infringement of these patents are decided by courts in accordance with patent law of that country.

Patenting of life forms or biological material is called biopatenting and such a patent is called biopatent. Before, 1980, life forms could not be patended and the discovery of oil-eating bacterium (genetically engineered Pseudomonas) by a non-resident indian scientist Dr. Chakraborty (called 'super bug', used for cleaning up oil spills) was first to be patended under standard US patent (i.e., first biopatent). genetically engineered mouse called 'oncomouse' (carrying a human cancer gene) was protected by a US patent in 1988 and this was another milestone in patenting of life forms or biopatents. Issue of biopatents has also been recently discussed in india.

In plant biotechnology, live plants, naturally occuring  microbes, micro propagation, tissue and organ culture techniques, biological controlo pests or hybrid varieties cannot be protected using patents. New crop varieties cannot be patended but transgenic animals and plants can be patended.

The genes and DNA sequences which normally fulfill the requirement of patents have no difficulty in getting patent protection. In 1991, the National Institute of Health (NIH). USA, applied to patent some DNA sequences useful in human gene mapping. This application was opposed on the grounds that it will slow down  the flow of scientific information and stop international co-operation and it failed because of cost.  A second application on DNA patenting by a private company was turned down on the grounds that the research has been done with public money. But in 1997, th eUS patent office agreed to allow patenting of gene strands with no biological function  but likely to have indutrial use. The result is that big academic labs and companies have been sequencing and patenting sequences in order tosecure private investment. In order to distinguish  DNA sequences from their natural counterparts (which cannot be patended), a patent application must state tht the invention has been purified or isolated or is part of recombinant  molecule or vector. Some pharmaceutical companies and NIH (National Institute of Health ) are compiling public databases of sequences toensure public acces. Further donor content is a concern in human DNA patenting .

Terminator technology for seed protection:
A patent under the title'control of plant Gene Expression' was issued on March 3, 1998 jointly to delta and pineland (D and PL) and the US department of Agriculture (USDA). A canada based NGO, RAFI (Rural Advancement Foundation International) dubbed the technology as 'Terminator technology', as they beleived that it will be used to ensure that farmers may not be able to save the seeds from their harvest for replanation. The inventors of this technology have already applied for the patents in atleast 78 countries. Indian governemnt has banned the entry of any seed material into the country that may carry terminator gene and has decided not to grant patent to D and PL for terminator technology.

Patended method for terminator technology is based on a gene that produces a protein that is toxic to plant and thus does not allow the seed to germinate (RIP gene).

Arther Dunkel's Suggestion (1991, 92): Arther Dunkel's then director General, GATT ( General Agreement of Tariff's and Trade ) gave some suggestions for india, which include:

i. India will have to patent laws by 2003 to finally open the agriculture sector for patended biotechnology

ii. India will not be able to give any kind of subsidies for production of oil seeds or pulses as the international prices of these commodities  are lower than domestic price.

iii. India  will have to introduce product patents and to increase in duration of patents.

iv. India will have to open its markets to foreign patent holders.

Biotechnology - Major application

1.Monoclonal antibodies: Antibodies are the specialized proteins, which are produced inside the host body in response  to foreign substances called antigens (mostly protein or polysaccharides) and provides immunity to the host against antigens. Antibodies are produced in Lymph nodes, spleen and liver.

Monoclonal antibodies (Mabs) are the specialized antibodies, which are specific to only one type of antigens. The concept of monoclonal antobodies was put forward by georges Kohler and Cesal Milstein in 1974 and these antibodies are produced outside the body by hybrid cell culture technique, known as Hybridoma technology. For this technology, Kohler and Milstein were awarded Nobel Prize in 1984.

In this technique, the antigens (against which antibodies are needed ) are injected into rat is removed and from spleen, some lymphocytes (which are responsible for antibody production) are isolated. These lymphocytes are mixed  with myeloma cells (tumour cells isolated from cancer of bone marrow). Some of these lymphocytes  and myeloma cells fuse together to form hybridoma cells (by somatogamous fusion). These hybridoma cells have capacity of antibody production  (of lymphocytes) and rapid cell division (of cancer cells). These hybridoma cells in cultural conditions produce large quantities of specific and pure (monoclonal) antobodies, which are separated and used in cure of different diseases.


2. Vaccines: Production of antibodies against antigens, is the basis of immunity. Vaccine in fact is dilute dose (suspension) of antigens, which is used for developing artificial or acquired immunity . The process of inoculation of vaccine is known as vaccination  and Louis pasteur (1850) is responsible for understanding the basis of vaccination  and immunization. (Edward Jenner 1970, however made earlier studies in relation to small pox). A vaccine contains either weakened (polio, smallpox and measles vaccines)or even killed pathogens( typhoid vaccine ) which have still antigens to induce antibody production. In some cases (like botulism and tetanus), toxins produced by pathogens serve as vaccines . A lare number of vaccines (called first generation vaccines) against different viral and bacterial diseases have been produced.

In recent years, some new vaccines called second generations and third generations vaccines have been developed . Second generations vaccines are those, which are produced by recombinant DNA technology or genetic engineering . e.g., vaccine for herpes virus and Hepatitis B. Third generations vaccines are produced synthetically or are synthesized vaccines, e.g., for feline leukemia virus and foot  and mouth diseases  virus.

3. Amino acids : An important basic amino acid, lysine is produced from diaminopimelic acid. (constituent of cell wall of bacteria, e.g., E.coli by activity of bacterium Enterobacter


4. Some other food products : Cheese is produced by activity of different species of lactobacillus and Streptococcus.
Penicillium camembertii and P. roquefortii are used to give flavour to cheese (camembert cheese and roquefort cheese).
Similiarly, butter is produced from cream by activity of streptococcus lactis and Leuconostoc species.

5.Dextran : Sucrose by activity of Leuconostoc mesenteroides bacteria, gives rise to a complex polysaccharide called dextran, which is used in plasma tranfusions.

6. Cholesterol lowering statins: Monascus purpureus (red yeast ) is a species of mould that is purplish red in colour and is known by the names ang-khak rice mould, corn silage mould, maize silage mould, and rice kernel discoloration. This fungus is most  important because of its use in the form of red yeast rice and in the production of certain fermented foods in china. However, discovery of production of cholesterol  statins bythe mould has prompted research into its possible medical uses. It produces a number of statins. The naturally occuring lovastatins and analogues are monacolins K, L,J and also occur in their hydroxyl acid forms alongwith dehydroxymonacolin and compactin (mevastatin).The prescription drug lovastatin, identical to monacolin K, is the principal statin produced by Monascus purpureus.

7. Tissue cultue: This is also a latest method of crop improvement. Besides also used in manufacture of antibodies, alkaloids, and dyes (like shikonin, a red dye used in silk industry and in treatment of burns is obtained from culture of cells of lithospermum root)

8. Single cell proteins (SCP): This terms was first used in 1966. But this term is misleading now a days because it denotes not only the isolated cell protein but it is used for any microbial biomass from uni-multi-cellular bacteri, yeast, algae or filamentous fungi which can be used as food or food additives. In general, microbial biomass contains 45-55 % protein, though in some bacteria, upto 80% protein content present, Large scale production of microbial biomass (SCP) has advantages over traditional methods of producing proteins as microbes have high rate of multiplicationm, have high protein content, can utilize large number of different carbon sources (waste products also) and microbial biomass production is independent od seasonal and climatic variation. Most commonly used microbes are Spirulina (blue green alga), Chlorella (green alga), Saccharomycopsis) lipolytica (fungus), mushrooms, etc.

Methlophilus methylotrophus bacteria have been investigated for use in single cell protein production. These bacteria have a generation time of about 2 hours and mainly used in animal feed as bacteria, in general produce a more favourable protein composition than yeast or fungi. Therefore, the large quantities of SCP animal feed using bacteria. The resulting product appears pinkish white, is odourless, tasteless and non-toxic which consists of about 65-75% protein of balanced amino acids composition.

Probiotics are live microorganism thought to be beneficial to the host organism. This term has been derived from latin ('pro -for+ biotics, derived from noun 'Bios-Life). According to currently adopted definitions by FAO/WHO, probiotics are live microorganism which when administered in adequate amounts confer a health benefits onthe host. Lactic Acid Bacteria (LAB) and bifidobacteria are the most common types of microbes used as probiotics, but certain yeasts and bacilli may also be helpful. Probiotics are commonly consumed as part of fermented foods with specially added active live cultures; such as yoghurt, soyyohurt or as dietary supplements. At the start of 20th century, probiotics were thought to beneficially affect the host by improving its intestinal microbial balance, thus inhibiting pathogens and toxins producing bacteria.

9.Sewage treatment and methane production using microbial systems: Conventional sewage treatement involves the use of the naturally developing micro-organisms within the sewage treatement system. Now a days the sewage is onoculated with a specific micro organisms, specially selected for that particular sewage treatement process. These organisms are called ' Starter cultures'

A strain of Pseudomonas putida containing plasmids has been developed, which degrade octane, xylene, metaxylene and camphor.

Methane is produced during anaerobic decomposition of sewage and other organic wastes by bacterias. The methane is collected and used as fuel in many countries.

Biotechnology And Sustainable Agriculture

Green revolution and high impact agriculture have satisfied the food requirement of most of the human population as it has doubled since 1960. But increase in food productions has led to serious environmental problems, i.e., high impact agriculture is not sustainable for more than a few decades. So, more and more farmers are trying forms of agriculture that cause less environmental problems. Sustainable agriculture (or alternative agriculture of low-input agriculture) is based on beneficial biological processes and environment friendly chemicals. In sustainable agriculture, certain modern agricultural techniques are combined with traditional farming methods. Sustainable Agriculture may be simply defined as, "Agricultural methods that rely on beneficial biological processes and environment friendly chemicals rather than conventional agricultural  techniques."
In other way, Sustainable Agriculture may be defined as, "The successful management of resources for agriculture to satisfy human needs while maintaining or enhancing the natural resource base and avoiding environmental degradation".

Hence we can say, Sustainable Agriculture is not a single programme but there is a series of programmes that are adapted for specific soils, climates and farming requirements. Water and energy conservation, crop rotation, breeding of disease resistant crop plants, use of biofertilizers (BGA fertilizers mycorrhizae, symbiotic and asymbiotic nitrogen fixers, organic fertilizers, green manures), biological pest control methods and bio-pesticides or Integrated Pest Management (IPM) are practiced in sustainable agriculture.

(Bio-fertilizers and biological pest control are given separately in detail in this unit.
In addition to these practices, biotechnology also plays a promising role in sustainable agriculture. Biotechnology held the potential of marrying biology and engineering and is essentially interdisciplinary area consisting of biochemistry, molecular biology, microbiology and genetics. Plant biotechnology includes two major technologies cell, tissue and organ culture and the recombinant DNA technology/genetic engineering. Cell and tissue culture has many commercial applications which include micro propagation, haploid breeding, productions of secondary metabolites and use in production of transgenic crops or genetically modified crops. Recombinant DNA technology has two major uses namely molecular marker technology and production of genetically modified transgenic crops. In general genetic engineering promises greater productivity and more nutritious foods.

The molecular markers have been utilized in characterization, maintenance and sustainable use of PGRs, (plant genetic resources). Their main uses in plant breeding include selection of desirable plants in early segregating generations at the seedling stage and selection of diverse parents for hybridization.

The most important use of plant biotechnology has been in the production of superior transgenic crops which are having not only resistance to different biotic and abiotic stresses but are also having improved nutritional quality of food.

Using traditional breeding methods, it takes 15 years or more to introduce disease resistance genes into a particular crop plant but genetic engineering or recombinant DNA technology takes a fraction of that time in achieving the same goal. Further in this desirable genes from any organisation can be used.

Genetic engineering may produce more nutritious food plants because they would contain all the essential amino acids. Recently in 2000, an international team of scientists has successfully engineered rice grains to produce beta-carotene or vitamin A (Golden rice or miracle rice).This rice has the potential to improve world health because about half of the world's population use rice as their staple food and rice is a poor source of many vitamins including vitamin A.

Using genetic engineering,crop plants resistant to insect-pests, viral diseases, drought, heat, cold, herbicides and salty or acidic soils can be developed.

In a cross of agriculture and medicine, molecular biologists are developing GM plants ( Genetically modified plants) that give edible vaccines, e.g., antidiarrhea vaccine and cholera vaccine both contain in raw potato. Similarly, extract preventing tooth decay and a vaccine against ulcers have been obtained from GM crops. Bananas (more palatable than potato) are also being genetically modified to provide edible vaccines, Edible vaccines hold important role for developing countries as they can be grown where most needed and they will not require clean needles and constant refrigeration.

Genetically modified crops transgenic crops have already transformed agriculture and they have been grown in more than 100 million acres of farmer's fields, each in year 1999 and 2000 despite of protests and debates.

Plant genomics is the latest or new field of plant biotechnology. Under this discipline, complete genome of Arabidopsis has been sequenced and of rice is being sequenced. This will lead to find the functions of all genes (reverse genetics), which will be having a great impact in field of cytogenetics, plants breeding and plant pathology. The Knowledge from genomics research will be utilized in detection, isolation and manipulation of genes in a variety of crops and thus, novel transgenic crops will be produced. As there is a great complexity of plant system, so let us see which way the wind blows.

Thus plant biotechnology has a great potential to improve agriculture

Branches of biotechnology

Biotechnology has over many totally different branches that are referred to by dissimilar terms principally noticeable with dissimilar colours to clarify the biotechnological field that it's employed in. the bulk extensively used ones are introduced here. first of all there's the red biotechnology that's used for medical processes, like finding genetic cures by prying genomic manipulations and making organisms to provide antibiotics.


Green biotechnology is employed in orientation to agricultural method that use biotechnology.. Some samples of that might be the event of transgenic plants that ar calculated to survive beneath precise environmental circumstances. an oversized goal of the inexperienced biotechnology is to expand a lot of atmosphere friendly solutions, as an example to search out the simplest way eliminate the requirement for pesticides.

Green biotechnology that is a lot of sometimes called Plant Biotechnology could be a speedily increasing field among fashionable biotechnology. It basically involves the gap of foreign genes into inexpensively vital plant species, leading to crop improvement and also the production of novel product in plants.

The term used for producing biotechnology is white biotechnology. this kind of biotechnology is employed to decrease the prices for manufacturing industrial provides that occur once ancient processes ar used. as an example, white biotechnology will expand AN organism that's capable to provide an explicit helpful chemical by natural processes quite than by industrial ways in which it absolutely was done earlier.


The industrial biotechnology society sometimes accepts a casual divide between producing and pharmaceutical biotechnology. an example would be that of company rising plant life to provide antibiotics, e.g. antibiotic from the genus Penicillium fungi.

Some extra samples of branches of biotechnology ar blue biotechnologies that handle marine and a marine usage of biotechnology, however that's not terribly extensively used. once discussing regarding not the straight analysis a part of biotechnology then bioeconomy is employed to speak regarding the savings and also the economical edges that biotechnology brings.

Biotechnology with marine organisms, feed into cultivation, fauna and fish health, marine natural product (including medicines), biofilms, bioremediation, marine ecology and bio-oceanography and different marine product (e.g. enzymes)

Biotechnology in india

In india, biotechnology has gained momentum after 1980-85.The Indian Agricultural Research Institute (IARI), New delhi has taken  initiative in fundamental and applied aspects of molecular biology and biotechnology. Tissue culture technique and micropropagation have been  adopted for producing seedlings of horticulturally tested varieties of orchids and banana. Protoplast fusion and somatic hybridization have also been applied for producing the rare hybrids of ornaments and vegetable plants. Council of Scientific and Industrial Research (CSIR), New delhi and Indian Council of agricultural Research (ICAR) have taken keen interests towards fundamental studies in molecular biology (such as protoplast culture, tissue culture, organ culture, translation and transcription processes in prokaryotes, immobilization of plant cells, clonal propagation, etc.)

National Biotechnology Board(NBTB) has been set up for promoting research in biotechnology on the whole i.e., in microbial genetics, gene biotechnology, DNA sequencing, genome study, tissue culture, enzyme technology, fermentation technology, immunization technology, gene amplification and DNA finger-printing. In india DNA finger printing was first carried out Dr. Lal ji singh Director, centre for cellular and molecular  biology (CCMB), hyderabad. He was able to synthesize DNA probe by himself for carrying out DNA fingerprinting in india. This techique is applied for solving the complicated problems of murders (in absence of eye witness), rapes an dispute parentage.

United National Industrial Development organization (UNIDO) have taken initiatives in establishing an international  Centre for genetic Engineering and Biotechnology (ICGEB) in New delhi to provide tools and techniques for establishing modern biotechnology centres at different Universities and Institutions working in biotechnology and genetic engineering in various developing nations. Lal Bahadur shastri centre for biotechnology has recently been established at IARI, New Delhi  for basic and applied researches in non-gene and gene biotechnology in india.
Advantages of tissue culture in cell culture, organ culture, micropropagation, conservation of rare and endangered germ-plasts, protoplast culture and its use in somatic hybridization and advantages of somaclonal variations have been discussed in coming article of Plant breeding and Tissue culture. They are now widely used in pharmaceutical industries or fermentation based industries.

Gene Biotechnology

In Gene biotechnology, suitable techniques are developed to synthesis gene/s in vitro, either by chemical synthesis method (as was used by Dr. Har govind khurana for synthesizing first gene in vitro ) or by using DNA or RNA obtained from the organism themselves (i.e from natural source). It needs a well organized Molecular biology lab, with facilites of gel electrophoresis, southern blotting, autoradiography, recombinant DNA technology, gene synthesis, PCR technique, tissue culture technique, etc

Recombinant DNA technology has extensively been used for introducing genes of interferon, insulin, somatotropin, nif- genes, bt-genes, etc., in plasmid DNA obtained from suitable bacterial organisms. The recombinant DNA is then inserted into suitable host cell (bacterial cells used as cloning organism) for gene cloning (producing indentical copies of recombinant DNA). This technique has been applied for producing genetically modified or engineered bacteria carrying desired gene/s.)

Transgenic organisms or Genetically Modified (GM) organism (plants, animals or microbes) are the most valuable gifts of biotechnology as they carry desired genes. They are capable of fulfilling the food requirements of growing world populations. Recombinant DNA technology has now been used widely not only in industries or pharmaceuticals concerns but also in agriculture and environmental biology.
Non-gene biotechnology is infact dealing the modern aspects of microbiology. Microorganisms can show be exploited for producing antibiotics, antibodies and industrially important biochemicals including enzymes.It has opened the door for several industrial and agro based companies for biorevolution. Many Nobel Laureates have also offered their services to the biotechnological organizations carrying out researches in advanced microbiology and biotechnology.

In addition to these practices, biotechnology also plays a promising role in sustainable agriculture. Biotechnology held the potential of marrying biology and engineering and is essentially interdisciplinary area consisting of biochemistry, molecular biology, microbiology and genetics. Plant biotechnology includes two major technologies cell, tissue and organ culture and the recombinant DNA technology/genetic engineering. Cell and tissue culture has many commercial applications which include micro propagation, haploid breeding, productions of secondary metabolites and use in production of transgenic crops or genetically modified crops. Recombinant DNA technology has two major uses namely molecular marker technology and production of genetically modified transgenic crops. In general genetic engineering promises greater productivity and more nutritious foods.

The molecular markers have been utilized in characterization, maintenance and sustainable use of PGRs, (plant genetic resources). Their main uses in plant breeding include selection of desirable plants in early segregating generations at the seedling stage and selection of diverse parents for hybridization.

The most important use of plant biotechnology has been in the production of superior transgenic crops which are having not only resistance to different biotic and abiotic stresses but are also having improved nutritional quality of food.

Using traditional breeding methods, it takes 15 years or more to introduce disease resistance genes into a particular crop plant but genetic engineering or recombinant DNA technology takes a fraction of that time in achieving the same goal. Further in this desirable genes from any organisation can be used.

Genetic engineering may produce more nutritious food plants because they would contain all the essential amino acids. Recently in 2000, an international team of scientists has successfully engineered rice grains to produce beta-carotene or vitamin A (Golden rice or miracle rice).This rice has the potential to improve world health because about half of the world's population use rice as their staple food and rice is a poor source of many vitamins including vitamin A.

Using genetic engineering,crop plants resistant to insect-pests, viral diseases, drought, heat, cold, herbicides and salty or acidic soils can be developed.

Foods reformed by Biotech

In the last five years, the area under transgenic crops has increased enormously in USA, China and some other advanced countries. In USA, 35% of corn and 55% of all soybean planted were transgenic in 1999. As soybean and corn are used as ingredients in a wide range of food products, many foods currently contain ingredients  derived from genetically engineered or transgenic crops. The area under transgenic crops will continue to rise and thus will result in more food derived from transgenic crops. The foods derived from transgenic crops are called ' GM foods' or genetically modified foods.

Health and food safety concerns have been raised to ensure the safety of GM foods throughout the development and commercialization of transgenic crops. The main concerns  about the potential of GM foods/ crops relate to:

Increase in toxins
Introduction of allergens
Changes in levels of essential nutrients
Reduced efficacy of antobiotics
Issues of food safety from GM crops came with a 3-years study conducted by Dr. Arpad Pusztai at Rowett Research Institute, Scotland in 1999, who showed partial suppression of growth and of development of immune system in rats fed on GM potato.

Dr. March Lappe (1999) showed that certain GM foods (GM soybean ) have lower levels of vital nutrients especially phytoestrogen and isoflavones which protect the body from heart diseases and cancer.


Nordlee( 1996)  reported that Brazilnut (Bertholletica excelsa) genes in soya caused epileptic fits.similarly, Shrimp gene in strawberry induced allergic response.

During early course of development of trangenic crops, a concern was raised that transfer of antibiotic resistance transgene from plants  to pathogene in environment or in the gut of humans or animals would compromise antibiotic therapy by rendering pathogens immune to the effects of the antibiotic.

On the other hand genetically modified crops are being developed with enhanced nutritional qualities such as increased iron (Fe) or vitamin A content in edible plant parts (e.g., 'Golden rice' or 'Miracle rice' rich in vitamin A and iron). Iron fortification of rice  has been done by soybean ferritin gene.

The US food and drug Administration (USFDA) established extensive guidelines to address the concerns and ensure the safe commercial introduction  of GM crops for use in food and feed. These guidelines have been developed through evaluation of scientific data consultation with experts from around the world. Foods containing GM crops meeting the FDA guidelines are deemed to be as safe and nutritious as their non- GM counter parts already in market.

There have been debates in many countries on whether or not, it should be mandatory to label the genetically engineered food. In USA, no such mandatory lebelling has been considered necessary, unless the transgenic food or GM foods differs significantly in composition or is allergic, e.g., tomato having significantly reduced content of vitamin C should be lebelled. Similarly , tomato having a gene from peanut should disclose this as peanut contains allergens. Transgenic canola (oil seed rape ) having a gene of brazilnut (being allergenic) is considered to have no commercial utility and hence should be lebelled for consumers
GM Foods safety guidelines established using current scientific knowledge are not static and will continue to refined as new knowledge is gained in areas related to food safety .

Enzymes worn in biotechnology or r-DNA Technology

DNA ligase enzyme- A type of enzyme used to seal the cut ends of DNA  fragments. It is a type of molecular adhesive used during joining of desired gene with suitable vector DNA. DNA ligase was first discovered by Dr. Khorana from T4 bacteriophages and thus named as T4-ligase.
Endonuclease enzyme- It is a tiny biological scissor used to cut foreign DNA and plasmid DNA before they are allowed to form recombinant DNA. It occurs naturally in some bacterial cells and acts as chemical weapon against the invading bacteriophages. It cuts both the strands of double stranded DNA and occurs in solution form.
Most of the restriction endonuclease enzyme a specific palindromic sequence for producing cuts and they usually show sticky end type cutting. The existence of restriction enzyme in bacterial cells was first demonstrated by Arber. (1906) in Haemophilus influenzae and named as Hind-III .So far, over 475 types of restrictions endonuclease enzymes have isolated and identified.

Exonuclease enzyme: The enzyme which cut single strand of DNA, digesting base pairs on 5' to 3' end or at single stranded nick of a DNA strand.

Reverse transcriptase enzyme: It was discovered by Temin and Baltimore from retroviruses. Reverse transcriptase directs complementary DNA strand formation on the templates of m-RNA (RNA directed DNA synthesis or reverse of transcription process). It helps in in vitro synthesis of DNA or gene using specific m-RNA obtained from the cell. New gene can be obtained through this technique within a short duration using DNA polymerase enzyme and nucleotides of DNA molecules in triphosphate  form (dATP, dGTP, dTTp, dCTP.

S-I nuclease enzyme: It degenerates the single stranded DNA or single strand of a double stranded DNA with cohesive ends. As a result of action of S-I nuclease, cohesive ends are converted into blunt ends.

DNA polymerase enzyme: It is used for producing copies of DNA through PCR technique or replication  of DNA under usual  condition (DNA directed DNA synthesis). It was first discovered by Kornberg.

RNA polymerase enzyme: It is meant for transcription of DNA molecules, i.e, DNA directed RNA synthesis or synthesis of RNA on DNA templates. All types of RNA (m-RNA, t-RNA and r-RNA) are synthesized by DNA templates using RNA polymerase.

Crops regenerate by the help of Biotech

GENETICALLY  MODIFIED  CROPS  OR  TRANSGENIC  PLANTS /CROPS
The most important use of the  plants biotechnology  is  the production  of  superior  transgenic  crops/plants  which  are  having  not only  the  resistance  against a  variety  of  biotic  and abiotic  stresses, but also having some improved value added properties  like nutritional quality  of  food.Transgenic plants are those plants in which a  foreign  gene  has  been  introduced  and stably  integrated  into host DNA. A gene that  has transferred  using  the  tools  of  molecular biology is called  transgene. The.The transfer or introduction  of  a  foreign  gene  result  in  synthesis  of  desirable  traits like disease  resistance, insect  resistance, herbicide resistance, etc.

Transgenic  plants have now been  produced  in more  than 60 plants species (mostly in  dicotyledonous families and  in some monocotyledonous  families  also).The  first transgenic plants were  produced  in tobacco (Nicotiana tobacum).Some other higher plants where transgenic plants have been produced are petunia hybrid,Lycopersicum esculentum (tomato), solanum tuberosum (solanum) melongena (brinjal), Lactuca sativa (lettuca) ,Brassica napus (oilseed rape or canola), Glycine max (soybean), piscum sativum (pea), Beta vulgaris (sugar beet), vitis vinifera (grape),Asparagus. Secale cereale (rye), Triticum aestivum (wheat), Zea mays (maize), Avena sativa (oat) and a gymnosperm picea glaucoma (white spruce), etc.

Some of the transgenic plants have been released for commercial cultivation,e.g.,'Flavr savr' and Endless Summer' tomatoes, 'Freedom II' Squash, 'high lauric' rape and seed or canola, 'Roundup Ready' soybean. Similarly genetically engineered insect resistance potato (New leaf ), corn (Maximizer) and cotton (Boll gard), each having Bt toxin gene from B acillus thuringiensis, have been granted full registration. Transgenic plants/crops are also called genetically engineered crops or gm crops (genetically modified crops) or gmo (genetically modified organised)

Salt tolerant transgenic plants have been developed in Arabidopsis and tomato and heribicide resistance transgenic plants have been developed in maize, beet, rape, tomato, potato, tobacco, chicory and melon.

Specific gene transfer in plants is mostly done with Agrobacterium tumefaciens bacteria (causes crown gall) diseases in many dicots). The tumour causing ability resides in a plasmid called Ti plasmid of a tumefaciens. This plasmid has become important in plant breeding because specific genes can be inserted into the Ti plasmid by recombinant DNA technique and thereby changing the genotype and phenotype of the plant. A tumefaciens based gene transfer is hampered by host range  limitations.
               
Nematode resistance in tobacco plants has been brought about by RNA interference( RNAi) technique. In the technique, a complementary RNA binds to mRNA to form a dsRNA, which is unable to translate and hence the expression is blocked. Here a nematode specific gene is introduced in host plant. When this plant is infected by the nematode, the nematode specific gene is triggered and it produced a single stranded RNA which is complementary to RNA produced by the nematode is order to infect the host plant. This complementary RNA binds to the mRNA of the nematode to form a double stranded RNA that cannot be translated. Hence the nematode is unable to infect the transgenic tobacco plant.

Gene transfer can also be made directly without using biological vectors (plasmids, bacteriophages, etc.) Naked plant cells as protoplasts have been found ideal for vectorless gene transfer. Some vectorless gene transfer methods are:

a)Gene transfer using practicle bombardment or biolistics which involves directly shooting a piece of DNA into the recipient plants tissue by a gene gun. Tungsten or gold beads (which are smaller than the plants cells themselves) are coated in the gene of interest and fired through a stopping screen, accelerated by Helium, into the plants tissue. The particles pass through plant cell, leaving the DNA inside. This technique is being used in monocotyledonous and dicotyledonous plants successfully. It is relatively simple laboratory technique. The transformed tissue is selected using marker genes such as those that code for antibiotic resistance. Whole plants are then regenerated from the totipotent transformed cells in culture, containing a copy of transgene in every single cell.

b)technique
c)Macroinjection
d)Lipofection
e)Silicon carbide fibre-vortex
f)Ca-DNA co-precipitation method
g)Electroporation
h)Ultrasonication
i)UV-laser microbeam ( However, success in few systems).


Recently , transgenic plants have been explored for production of biologically active peptides and proteins (having pharmaceutical applications including use as vaccines immunomodulators , growth factors, hormones, blood proteins and enzymes).These proteins and peptides play a key role in production  of edible vaccines and antibodies , which are convenient and inexpensive for immunotherapy. An edible vaccine against  diarrhoea has been expressed in potato and against  dental caries in tobacco. Research on transgenic plant-based production  of various vaccines and antibodies is in early stages but these generate a hope towards low-cost edible vaccines and antibodies from plants.

Plant bio technologies in india have claimed the production of an edible vaccine against cholera in tomato. This tomato will be light bluish in colour and is expected to be in market in next years.

Golden rice or Miracle rice (rich in vitamin A or b-carotene and iron ) and decaffeinated coffee are also valuable achievements of gene transfer technology.

Biosafety of Transgenic plants: Nowadays, there is a debate on biosafety issues of transgenic plants. The key points are:
i. Potential of GM crops to become weeds.
ii. Opportunities of gene flow from a GM plant to other plant.
iii. Impact of growing transgenic plants on biodiversity.
iv. Capacity of pest and pathogens to adapt to the cultivation of GM crops.

Ignoring the risks involved in cultivation of transgenic crops, in China, 2300 hectares of land has been placed under transgenic crops . In india, however onlyone transgenic crop (bt-cotton) has so far been released for cultivation. Some srains of Bacillus thuringiensis produce proteins which have the property of killing certain insects. Specific by toxins genes isolated from Bacillus thuringiensis are incorporated into cotton and other crop plants. As most of the bt toxins are insect-group specific, so choice of genes depends upon the crop and the targeted pest. The bt toxin is coded by a gene called cry, Which are of different types like cry 1Ac, cry II Ab (Controlling cotton bollworms) and cry 1Ab (controlling corn borers). Recently in March 2002, Government of India has permitted the commercial cultivation of three varieties of bt-cooton, the first genetically modified crop of the country. This has been developed by MAHYCO ( Maharashtra Hybrid Seeds Company ) in collaboration with American company Monsanto. bt-cotton is resistant to Boll-worm disease of cotton.

Types of Biotechnology

The process of Developing and creating of technological applications that uses Biological systems or living organisms is named Biotechnology. counting on the tools and applications that area unit developed; it helps to switch the product or processes for a specific use. Biotechnology additionally employed in agriculture and food production and also in making ready medicines. It plays a serious role in development of pharmaceutical therapies. The word "Biotechnology" was given by Hungarian within the year 1919. In order, to boost the cultivation of plants and taming of animals wide selection of ideas of biotechnology is employed with prolonged procedures for modifying living organisms in step with human desires. Breeding programs like artificial choice and coupling area unit used.

Medical Biotechnology:

Medical Biotechnology helps in preventing human diseases. the employment of living cells and cell materials is named Medical Biotechnology. The living cells and cell materials serve the aim for analysis and additionally helps turn out company product and diagnostic product that eradicate human diseases. The medical analysis studies area unit conducted by the medical biotechnologists and these biotechnologists work for educational and industrial desires. The biotechnologists conduct varied experiments that a part of the analysis that helps to spot, treat various diseases and the way to stop diseases. The biotechnologists are terribly no-hit in conveyance varied techniques within the field of medical Biotechnology. The contributions towards developing medicine and vaccines are majorly contributed by the commercial biotechnologists. Techniques like environmental clean-up, market microorganism pesticides and insect- resistant crops area unit all introduced by the Biotechnologists within the field of Biotechnology. In straightforward words, Medical Biotechnology is that the study of micro-organisms like bacterium, plants and animals. It additionally facilitates in learning enzymes that is at hand in industrial and producing sectors and processes that help in manufacturing foodstuffs, necessary medicine used for creating medicines and artificial hormones.

The analysis studies associated with desoxyribonucleic acid (DNA) is that the pioneer illustration of the inventions within the field of medical biotechnology additionally as well as the invention of internal secretion and somatotrophic hormone. The study of gene-splicing involves in distinctive the human genes and this study is completed by several biotechnologists within the field of Biotechnology. Diseases like Parkinson’s and Alzheimer’s syndrome is presumably area unit cured with the assistance gene-splicing.

Industrial biotechnologists work for personal firms for a specific style of analysis and that they area unit concerned in finding a specific medical issue or flaws within the analysis. Their findings area unit usually confined to invention of latest medical treatments and invention of medical medicine. The personal firms rent industrial biotechnologists to fulfill their money desires and goals of their business. the commercial biotechnologists usually prove and make a case for their findings to non-scientists; since the commercial biotechnologists lack higher cognitive process powers. Whereas, tutorial Biotechnologists usually work for non profit organizations, colleges, pioneer universities and government organizations like government hospitals and labs. Most of the tutorial Biotechnologists begin their study and analysis since their graduation days.

Agricultural Biotechnology

The technique used for improvising the plants, small organisms and animals is named Agricultural Biotechnology. DNA tests facilitate the scientists to live and develop solutions to boost agricultural productivity. It additionally helps to boost breeders’ so as to form enhancements in new crops and placental mammal. DNA plays a key role for brand spanking new enhancements and developments in Agricultural Biotechnology.

Agricultural Biotechnology is majorly employed in gene-splicing, Molecular manufacturers, Molecular medical specialty, Vaccines and tissue culture. for many years, the farmers are operating laborious to boost the standard of crops, plants and animals through choice and breeding processes. Since the 20th century method|the method} of breeding in agricultural biotechnology became a really at hand and a significant process for the farmers and within the agricultural business. the method of breeding helps to enhances flavour, it's gadfly resistant and it helps to extend the productivity of the endth results post agricultural method. The Breeding traits area unit passed from one generation to a different generation that facilitate of genes that is created out of the DNA tests. in step with the recent study the scientists have learned to figure in accordance to the genes or the DNA that is answerable for the long run dynamic  traits within the agricultural business. The cells those area unit gift altogether the living things as well as fruits, vegetables, and meat operate in accordance to the genes found within the cells gift in living things and eatables.

The use of contemporary biotechnology is incredibly necessary so as to boost the standard of crops in Crop production. Few genes area unit transferred from one crop to a different to boost the standard of every increased  crop cultivated. This method of transferring the genes helps to boost the organic process qualities, style {and the|and therefore the|and additionally the} look and feel of the eatables it also reduces the dependency of on artificial fertilizers and pesticides and agrochemical product for the cultivation of those crops.

The process of modifying the living organisms is named Animal Biotechnology. it's employed in science and engineering of Animals. the most aim and goal of Animal biotechnology is to supply quality product and enhance small organisms for a specific agricultural uses. samples of Animal biotechnology area unit new breeds of animals introduced by humans and with the assistance of biological research method (introducing identical animals). the normal Breeding techniques area unit employed in animal biotechnology decades agone.

Industrial Biotechnology:

The use of Plants, marine organisms, algae, fungi and small organisms is named industrial Biotechnology. Biological resources area unit accustomed turn out the chemicals, materials and energy needed for the event of commercial merchandise. Industrial Biotechnology is additionally well-known by the name “white Biotechnology”. it's additionally accustomed build Biobased product in sure sectors like paper, pulp, textiles, biofuels and biogas. Industrial Biotechnology helps the environmental edges and it additionally helps in up the performance of the business and a worth augment its product too. Industrial biotechnology has tested to be the foremost tributary, innovative and promising approaches towards the commercial growth and it's helped in lowering gas emissions. it's additionally contributed the impacts in climatical modification within the industrial sector and within the alternative sectors too. because the industrial technology improvises day by day, generations once generations it helps not solely the changes within the weather conditions however additionally it helps area unit economy to growth within the industrial sector and within the alternative sectors too. because the business improvises there's growth and scope for innovation and technology. It additionally helps in pollution interference, resource conservation and price reduction. If correct measures area unit planned, dead and experienced industrial biotechnology is tested the most important sector as compared to Agricultural and medical biotechnologies or sectors. Industrial biotechnology is stated because the Third wave in Biotechnology. in step with the newest study and update new industrial processes is thought of from the economic science laboratory study to commercial sectors or applications 2 to 5 years down the road.

Industrial Biotechnology helps in protective the setting whereas it offers the business business in cut techniques and helps to make additional and additional markets within the business world. additionally it's not a time intense method as compared to drug product endure. it's thought of because the most fastest and also the best business for the companies and for the varied markets. Since the invention of commercial biotechnology is thus new that blessings and advantages of this sector is unknown and not understood and explored by the customers and business itself. The enhancements within the industrial processes has improved such a lot that sure inventions that have happened thus far within the industrial biotechnology sector has helped to up-lift the business to a larger height within the world. it's not solely reworking the producing method however additionally serving to North American country to create to newer product that we tend to may have did not imagine a few years agone. it's helped in preventing air and pollution this can be one in all the most effective illustrations of business Biotechnology. alternative illustration of commercial biotechnology includes the assembly of soured product like brewage, Cheese, food and vinegar. As time passes by invention during this sector can keep improvising and by overcoming the DE-merits it'll reach larger heights.

Valuable success in the field of Biotechnology and genetic Engineering

Biotechnology is a recently developed  technology  concerned  with application  of living cells or tissues or their components for the benefits of mankind  and thus it is a package  term- for Biological technology (-a technology based on application of living organisms). It is a multidisciplinary approach  using knowledge of microbiology, tissue, culture, genetic engineering and chemical  engineering. It is a most vibrant discipline of modern molecular genetics.

The science of biotechnology is only 2-3 decades old but its contribution in meidcal sciences, agriculture, environmental protection and pollution abatement are noteworthy. It has helped in  in vitro synthesis of human insulin, interferon vaccines, monoclonal antibodies, antibiotics of latest generation and purification of drugs for cure of human diseases. It has produced high yielding plants, genetically modified or transgenic plants (plants carrying suitable foreign genes) and disease resistant plants for various agricultural operations. Genetically modified plants have shown their capacity of nitrogen fixation (when nif- genes were inserted), insect repelling capacity (when bt- genes were introduced in cottons) and vitamin synthesizing ability (when vitamin A gene was inserted in paddy). Superbag has been created using plasmids of many varieties of Pseudomonas putida to deal with oil solution problem in ocean due to leakage in oil tankers.

1952  Hersey and Chase                 First reported that the infective part of bacteriophages is their DNA

1957 Isaacs and Lindeman              First isolated interferon as an anti viral substance

1958 Lederberg                               First reported genetic recombination in bacteria using E.coli

1958 Sanger                                    First worked out the sequence if amino acids

1960 Arber                                      First reported presence of restrictions Endonuclease in cells of bacteria

1969 Har govind Khorana              First discovered  DNA ligase in T4 - bacteriophase

1970 Har govind Khorana               First synthesized gene in vitro.It was gene for Alanine t-RNA of yeast                                                           having 77 base pairs. Later on he synthesized a gene of tyrosine t-                                                                 RNA if E.coli which was biologically functional for suppressor t-RNA

1970   Harris                                    First discovered DNA-RNA hybridization technique for synthesis of                                                            gene in vitro         

1970 Hammilton  Smith                  Isolated restriction endonuclease from Haemophilus influenza and

                                                           named Hind-III. He got nobel prize in 1978.

1971 Nathans                                   First used restriction to cut DNA molecule of monkey virus-II into 11                                                            pieces for preparing SV-40 gene map.

1973 Cesar Milstein                        First cultured myeloma cells (bone marrow cancer cells)

1974  Smith and Nathan                  First used restrictions endonuclease to DNA from specific                                                                             sites (palindrome)

1975 Temin and Baltimore             First discovered reverse transcriptase enzyme in sarcoma in sarcoma

                                                         virus (-a retrovirus )associated with cancer formation.

1977  Itukura et al.                         First chemically synthesized the DNA sequence of A and B chains of

                                                        insulin hormone and successfully inserted in PBR-322 plasmid of E.coli

1977 Bolivar and Rodriguez           First developed PBR-322 vector which was of 4.36 kb long and                                                                  double stranded.

1978   Arber, smith and nathans First discovered use of restriction endo- nuclease enzyme in genetic                                                               engineering or r-DNA technology.

1982  Brenner   First used plasmids in r-DNA technology

1982  Sanger   Got nobel prize for his outstanding contributions in genetic engineering.

1984  Niel, George kohler and Milstein Discovered Monoclonal antibodies producing technique using                                                                       hybridoma technology

1990  Murry, Joseph and Thomas Successfully disocovered about organ and tissue transplantation                                                                   technique and their application.

1993  Roberts Richards and sharp philip Discovered split gene and got Nobel prize

1994  Jeffreys    Discovered DNA fingerprinting  technique.

Concerning Genetic Engineering

Antenatal diagnose: Over 2500 genetic defects are known in human being due to certain defects in genes and the individuals carrying such defects are not normal . Their proper diagnosis at embryonal level is thus desirable using amniotic fluid which contain fetal DNA. It is called Antenatal diagnosis.

Antisense transcription : synthesis of m-RNA  occurs on sense strand of DNA. If is occurs on complementary strand of sense strand, it is called antisense transcription .Some rare horticultural plants have been developed through this technique carrying unique characters not found elsewhere.

Annealing - The process of denaturing ds-DNA by heat treatment and then slow cooling to allow the formation of DNA-RNA hybrid using desired RNA.c- DNA library - Complementary -DNA library produced by using template of m-RNA synthesizing  various types of c- DNA of a genome.

Biolipistics: A type of red dye synthesized by biotechnology and named as Shikonin. It is a naphthoquinone and it is produced  on mass  scale for commercial use in cosmetics using cell culture of Lithospermum erythrorhizon.

Blotting technique: Transfer of macromolecules (usually DNA fragments) by capillary action from a gel  plate to a nylon membrane (-a type similiar to zeroxing of printed sheet.)

Southern Blotting Technique (discovered by E.M southern) is used in DNA-RNA hybridization . Transfer of DNA strands ( fragments) after gel electrophoresis is done using nitrocellulose filter to facilitate further processing  when needed. It is used mainly in DNA finger printing technique. Northern Blotting TEchnique is applied for separation and transfer of RNA fragments from agrosol gel to nylon mesh. It is infact reversal of southern Blotting. In Western Blotting Technique analysis of protein is done. Protein fragments are electrophoresed in polyacrylamide gel and then transferred on to nitrocellulose  nylon membrane and the protein bonds are detected by using interaction with antibodies.

Callus: undifferentiated mass of cells produced in tissue culture experiment or in protoplast culture. Transgenic plants (genetically modified plants) can also developed using callus developed by the cells carrying recombinant DNA.

Chimera: A cell having genetically different DNA strands, developed due to addition of foreign gene into plasmid DNA to construct recombinant DNA.

Chimeric DNA: Hybrid DNA such as r-DNA, cosmid DNA, plasmid DNA, etc. r-DNA is produced by combination of vector DNA and desired gene. Cosmid DNA is produced by hybridization of plasmid DNA with a piece of lambda phage DNA including its cos-site. Phasmid DNA is produced by hybridization of lambda phage DNA with plasmid DNA where att-site of phage DNA is included.
Plasmid DNA +Foreign gene= r-DNA.
Cos-site containing phage DNA + Plasmid DNA= Plasmid

Clones : A group of genetically identical cells or organisms asexually descendant from a common ancestor (identical copies). all the cells or organisms of a clone have same genetic set up and they are thus representing the exact copies of the original.

cloning vectors:  self replicating entity to which the foreign DNA is covalently  attached for purpose of replication in host cell.

Cloning organisms: A host cell which can easily accept the donor gene. It contains its own genes and inserted genes and allowing them  to replicate in usual manner. Common cloning organisms used in biotechnology is E.coli.


Gene Therapy: Replacement of abnormal genes with normal genes through r-DNA technology . The first trial of actual (clinical) gene therapy was conducted in 1990 in U.S.A .  A four years old girl suffering with ADA (Adenosine Deaminase Deficiency) a lethal disorder was transfected with lymphocytes bearing the ADA gene carried by retroviral vector.

Monoclonal antibodies: Homogeneous immunization reagent synthesized technology for producing vaccines and for diagnosis of blood group and many genetic diseases.

Nif-genes: Nitrogen fixing genes, represented by a cluster of 17 genes and found in the cells of nitrogen fixing bacteria or in some cyanobacteria(e.g. Rhizobium, Klebsiella, Nostoc, Anabaema, etc. nif-genes are used in plant biotechnology .

pBR -322- It is constructed from recombination of two plasmid DNA (PBR- 318 and PBR-320). It consists ORI- site (site for origin of replication), R-site (gene for resistance against ampicillin  and tetramycin ) and unique site about  twenty types of restriction enzymes. It is thus a chimeric plasmid DNA. It is one of the standard cloning  vectors widely used in gene cloning experiments (derived from E.coli. plasmid  CoIE1), which is 4,362 bp DNA and was derived by several alterations in earlier cloning vectors. pBR-322 was named after Bolivar and Rodriguez, Mexican postdoctoral researchers who constructed it in 1977 in the laboratory of Herbert Boyer at the University of California San Francisco. It has genes for resistance against two antibodies tetracycline and ampicillin.

PCR technique (Polymerase chain reaction technique): A technique used for amplification of gene or DNA molecules .DNA strands are first denatured and then the two strands get separated to synthesize their complementary strands in presence of DNA polymerase enzyme using dATP, dCTP, dGTP and dTTP nucleotides as raw materials. This process is repeated several times to obtained desired amount of DNA for further  processing. PCR is discovered  by Kary Mullis in 1985 and is carried out in vitro. It mainly requires (a) the desired gene or segment of DNA to be amplified (b) nucleotide primers each about 20 bases long, (c) monomers of DNA as deoxynucleoside triphosphates such as dATP, dCTP, dGTP and dTTP. (d) heat stable DNA polymerase, viz., Tag polymerase(obtained from a bacterium,(Thermus aquaticus), pfu polymerase (obtained from a bacterium , (Pyrococcus furiosus) and vent polymerase (obtained from bacterium Thermococcus litoralis). pfu and vent Polymerases are found to be more efficient as compared to Taq polymerase.All these  polymerase are obtained from thermophilic bacteria .All these polymerase ar obtained from thermophilic bacteria and they are capable of showing their activities at a very high temp. like 75 degree to 80 degree C.
DNA is denatured at about 90 to 95 degree C. and then these 3 types of polymerases are used for amplification process. One DNA strand is capable of producing million of their identical copies through  PCR technique .

Plasmids : Extra- nuclear genetic material found in most of the bacterial cells as double stranded circular rings in addition to chromosomal DNA. They have proved to be the best vector for carrying desired gene/s for recombinant DNA technology. Plasmids can be  easily taken out from the bacterial cells through  diffusion by increasing the permeability of bacterial membrane and cell wall and can be reinserted (after constructing recombinant DNA ) into the bacterial cell for cloning.

Ri Plasmids: A plasmids of agrobacterium rhizogenes  which induces quick rooting in host organisms after infection.

Ti Plasmids: Plasmids of Agrobacterium  tumefaciens, capable ofproducing tumour in infected host cells through synthesizing cytokinin like growth hormone.

Somaclonal variations : Variations generated in somatic cells or tissues during prolonged storage in culture medium due to spontaneous mutation or due to action of chemicals used as ingredient of culture medium or due to change in position of transpozons.

Super bag: A genetically engineered variety of Pseudomonas putida carrying plasmids to hydrolyse xylane, nephthalene, camphor or toluene like  hydrocarbons found  in crude petroleum. It was first created by Dr. Anand mohan Chakrabarthy to deal with the problem of oil leakage from oil tankers in oceans .

Terminator technology : A technology used in producing plants of genetically modified high yeilding varieties developed recombinant DNA technology where seeds do not develop viable embryo and thus remain unable to germinate  and produce seedlings.


Transduction : Phage mediated transfer of genetic material from one bacterium to the other (as was first reported in Salmonella bacterium by Zinder and Lederberg).

Caulimo-virus or cauliflower Mosaic Virus (CaMV) is also used as a vector in plant biotechnology . It has double stranded and linear DNA of nearly 8 kb length. It causes diseases in wide range of dicot plants. Its replication involves reverse transcriptase enzymes similar to vetro-viruses .

Cybrids: A virus hybrids produced by somatic hybridization using protoplasts of two different parents. They carry cytoplasmic genes of both the parents but genome of only one parent.

Coliphage : A virus parasitizing on coliform bacteria or E.coli (e.g, )bacteriophage which posseses single stranded and circular DNA

Cybridization : Technique of producing cybrids by protocol plasmic fusion using Poly Ethylene Glycol (PEG) or electrofusion .

DNA finger printing : A technique in which DNA is cut into fragments of varying sizes using restriction enzyme and then fragments are separated through  gel-electrophoresis (DNA) fragments carry negative charges and thus they move towards the positive pole through the medium of suitable electrolyte. DNA fragments are then transferred to nylon membrane using. Southern blotting technique. Using radioactive DNA probe (synthetic DNA strands carrying radioactive bases), their presence is recorded on X-ray plate through autoradiography . These bands represent DNA fingerprints . This technique is used I forensic  science for solving complicated cases of murders and rapes (in absence of eye witness ) in or solving disputes in parentage .

DNA probe :Synthetic radioactive fragments of DNA used in locating the position  of specific  DNA fragments obtaining during  DNA  profiling of DNA fingerprinting after electrophoresis and southern blotting.

Electrophoresis : A technique of separating  DNA fragments of gel plate under the influence of electric current through the medium of suitable electrolyte.

Engineered bacterium: A bacterium carrying desired foreign gene/s for obtaining  specific products or for introducing desired gene/s  into organisms for producing genetically modified organism of desired nature.

Foreign DNA :DNA used for contracting recombinant DNA using suitable vector DNA or vehicle DNA. Foreign genes are also called passenger DNA and vector DNA is called vehicle DNA.

Gene back : Collection of genes of a genome with their correct identity  for future use in recombinant DNA technology or in formation of desired transgenic organism.

Gene Counseling: Gene counseling is done after knowing the correct family history and genetic diseases. If any in the family to work out possibility of future pregnancy on the basis of antenatal diagnosis .

Gene splicing : Cutting and rejoining the DNA sequence for r-DNA technology.

ELISA (Enzyme linked Immunosorbent Assay) Test: Used in case of detecting AIDS. It is useful in estimating the amount of specific antibody present in the antiserum used for the reaction.

Hybridomas: A hybrid of B-lymphocyte cell and myeloma cell (bone cancer cell) produced by somatic hybridization technique to synthesize desired Monoclonal Antibodies (MAB).

In vitro studies: Study outside the living body, i.e. by culturing the organisms in artificial medium or synthetic medium( as in case of tissue culture, organ culture, etc.)

In Vivo studies: Study within the living system or living body as in case of study of microsporogenesis in anthers, megaporogenesis in developing ovules or embryogenesis in fertilized ovules.