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Biotechnology (commonly abbreviated as biotech) is the broad area of biology involving living systems and organisms to develop or make products, or 'any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify product or processes for specific use' (UN Convention on Biological Diversity, Art. 2).[1] Depending on the tools and applications, it often overlaps with the (related) fields of molecular biology, bio-engineering, biomedical engineering, biomanufacturing, molecular engineering, etc.

Biotechnology Introduction O ne of the newest, yet controversial fields in science today is biotechnology. Biotechnology began in the 1970s after the development of genetic engineering that allowed scientists to m odify the genetic material of living cells. Genetic engineer ing is the manipulation of DNA.

For thousands of years, humankind has used biotechnology in agriculture, food production, and medicine.[2] The term is largely believed to have been coined in 1919 by Hungarian engineerKároly Ereky. In the late 20th and early 21st centuries, biotechnology has expanded to include new and diverse sciences such as genomics, recombinant gene techniques, applied immunology, and development of pharmaceutical therapies and diagnostic tests.[2]

  • 3Examples

Definitions[edit]

The wide concept of 'biotech' or 'biotechnology' encompasses a wide range of procedures for modifying living organisms according to human purposes, going back to domestication of animals, cultivation of the plants, and 'improvements' to these through breeding programs that employ artificial selection and hybridization. Modern usage also includes genetic engineering as well as cell and tissue culture technologies. The American Chemical Society defines biotechnology as the application of biological organisms, systems, or processes by various industries to learning about the science of life and the improvement of the value of materials and organisms such as pharmaceuticals, crops, and livestock.[3] Per the European Federation of Biotechnology, biotechnology is the integration of natural science and organisms, cells, parts thereof, and molecular analogues for products and services.[4] Biotechnology is based on the basicbiological sciences (e.g. molecular biology, biochemistry, cell biology, embryology, genetics, microbiology) and conversely provides methods to support and perform basic research in biology.

Biotechnology is the research and development in the laboratory using bioinformatics for exploration, extraction, exploitation and production from any living organisms and any source of biomass by means of biochemical engineering where high value-added products could be planned (reproduced by biosynthesis, for example), forecasted, formulated, developed, manufactured, and marketed for the purpose of sustainable operations (for the return from bottomless initial investment on R & D) and gaining durable patents rights (for exclusives rights for sales, and prior to this to receive national and international approval from the results on animal experiment and human experiment, especially on the pharmaceutical branch of biotechnology to prevent any undetected side-effects or safety concerns by using the products).[5][6][7] The utilization of biological processes, organisms or systems to produce products that are anticipated to improve human lives is termed biotechnology.[8]

By contrast, bioengineering is generally thought of as a related field that more heavily emphasizes higher systems approaches (not necessarily the altering or using of biological materials directly) for interfacing with and utilizing living things. Bioengineering is the application of the principles of engineering and natural sciences to tissues, cells and molecules. This can be considered as the use of knowledge from working with and manipulating biology to achieve a result that can improve functions in plants and animals.[9] Relatedly, biomedical engineering is an overlapping field that often draws upon and applies biotechnology (by various definitions), especially in certain sub-fields of biomedical or chemical engineering such as tissue engineering, biopharmaceutical engineering, and genetic engineering.

History[edit]

Brewing was an early application of biotechnology

Although not normally what first comes to mind, many forms of human-derived agriculture clearly fit the broad definition of 'utilizing a biotechnological system to make products'. Indeed, the cultivation of plants may be viewed as the earliest biotechnological enterprise.

Agriculture has been theorized to have become the dominant way of producing food since the Neolithic Revolution. Through early biotechnology, the earliest farmers selected and bred the best suited crops, having the highest yields, to produce enough food to support a growing population. As crops and fields became increasingly large and difficult to maintain, it was discovered that specific organisms and their by-products could effectively fertilize, restore nitrogen, and control pests. Throughout the history of agriculture, farmers have inadvertently altered the genetics of their crops through introducing them to new environments and breeding them with other plants — one of the first forms of biotechnology.

These processes also were included in early fermentation of beer.[10] These processes were introduced in early Mesopotamia, Egypt, China and India, and still use the same basic biological methods. In brewing, malted grains (containing enzymes) convert starch from grains into sugar and then adding specific yeasts to produce beer. In this process, carbohydrates in the grains broke down into alcohols, such as ethanol. Later, other cultures produced the process of lactic acid fermentation, which produced other preserved foods, such as soy sauce. Fermentation was also used in this time period to produce leavened bread. Although the process of fermentation was not fully understood until Louis Pasteur's work in 1857, it is still the first use of biotechnology to convert a food source into another form.

Before the time of Charles Darwin's work and life, animal and plant scientists had already used selective breeding. Darwin added to that body of work with his scientific observations about the ability of science to change species. These accounts contributed to Darwin's theory of natural selection.[11]

For thousands of years, humans have used selective breeding to improve production of crops and livestock to use them for food. In selective breeding, organisms with desirable characteristics are mated to produce offspring with the same characteristics. For example, this technique was used with corn to produce the largest and sweetest crops.[12]

In the early twentieth century scientists gained a greater understanding of microbiology and explored ways of manufacturing specific products. In 1917, Chaim Weizmann first used a pure microbiological culture in an industrial process, that of manufacturing corn starch using Clostridium acetobutylicum, to produce acetone, which the United Kingdom desperately needed to manufacture explosives during World War I.[13]

Biotechnology has also led to the development of antibiotics. In 1928, Alexander Fleming discovered the mold Penicillium. His work led to the purification of the antibiotic compound formed by the mold by Howard Florey, Ernst Boris Chain and Norman Heatley – to form what we today know as penicillin. In 1940, penicillin became available for medicinal use to treat bacterial infections in humans.[12]

The field of modern biotechnology is generally thought of as having been born in 1971 when Paul Berg's (Stanford) experiments in gene splicing had early success. Herbert W. Boyer (Univ. Calif. at San Francisco) and Stanley N. Cohen (Stanford) significantly advanced the new technology in 1972 by transferring genetic material into a bacterium, such that the imported material would be reproduced. The commercial viability of a biotechnology industry was significantly expanded on June 16, 1980, when the United States Supreme Court ruled that a genetically modifiedmicroorganism could be patented in the case of Diamond v. Chakrabarty.[14] Indian-born Ananda Chakrabarty, working for General Electric, had modified a bacterium (of the genus Pseudomonas) capable of breaking down crude oil, which he proposed to use in treating oil spills. (Chakrabarty's work did not involve gene manipulation but rather the transfer of entire organelles between strains of the Pseudomonas bacterium.

On the Performance dialog, click the Advanced tab and then click Change under the Virtual Memory heading. Now you’ll see the Virtual Memory settings as shown below. In Windows 7 and higher, the Automatically manage paging file size for all drives box is checked by default. If you’re running Windows 8 on a solid state hard drive with a Core i3, i5 or i7 processor, then you probably don’t. Jun 15, 2010  How to increase Virtual Memory size If your computer is slower then one of the most vital causes is the installed RAM. If your RAM size is small and you are running more applications at a time, then you may face different problems. Increase virtual memory windows 7. For example, your machine has 4GB memory and you are running multiple applications while it need a total 4.5 GB of memory so it will use the free space on the hard drive. It balances the size of the data according the empty space in RAM and transfer the rest to the hard drive. See Also: How to Increase Virtual Memory in Windows 10: A Quick. Aug 31, 2016  Windows 7 can run on a PC with 1 gigabyte (GB) of RAM, but it runs better with 2 GB. For optimal performance, boost that to 3 GB or more. Another option is to boost the amount of memory by using Windows‌ ReadyBoost. This feature allows you to use the storage space on some removable media devices, such as USB flash drives, to speed up your.

The MOSFET (metal-oxide-semiconductor field-effect transistor) was invented by Mohamed M. Atalla and Dawon Kahng in 1959.[15]Biosensor MOSFETs (BioFETs) were later developed, and they have since been widely used to measure physical, chemical, biological and environmental parameters.[16] The first BioFET was the ion-sensitive field-effect transistor (ISFET), invented by Piet Bergveld in 1970.[17] It is a special type of MOSFET,[16] where the metal gate is replaced by an ion-sensitive membrane, electrolyte solution and reference electrode.[18] The ISFET is widely used in biomedical applications, such as the detection of DNA hybridization, biomarker detection from blood, antibody detection, glucose measurement, pH sensing, and genetic technology.[18] By the mid-1980s, other BioFETs had been developed, including the gas sensor FET (GASFET), pressure sensor FET (PRESSFET), chemical field-effect transistor (ChemFET), reference ISFET (REFET), enzyme-modified FET (ENFET) and immunologically modified FET (IMFET).[16] By the early 2000s, BioFETs such as the DNA field-effect transistor (DNAFET), gene-modified FET (GenFET) and cell-potential BioFET (CPFET) had been developed.[18]

Revenue in the industry was expected to grow by 12.9% in 2008.

A factor influencing the biotechnology sector's success is improved intellectual property rights legislation—and enforcement—worldwide, as well as strengthened demand for medical and pharmaceutical products to cope with an ageing, and ailing, U.S. population.[19]

Rising demand for biofuels is expected to be good news for the biotechnology sector, with the Department of Energy estimating ethanol usage could reduce U.S. petroleum-derived fuel consumption by up to 30% by 2030. The biotechnology sector has allowed the U.S. farming industry to rapidly increase its supply of corn and soybeans—the main inputs into biofuels—by developing genetically modified seeds that resist pests and drought. By increasing farm productivity, biotechnology boosts biofuel production.[20]

Examples[edit]

A rose plant that began as cells grown in a tissue culture

Biotechnology has applications in four major industrial areas, including health care (medical), crop production and agriculture, non-food (industrial) uses of crops and other products (e.g. biodegradable plastics, vegetable oil, biofuels, and environmental uses).

For example, one application of biotechnology is the directed use of microorganisms for the manufacture of organic products (examples include beer and milk products). Another example is using naturally present bacteria by the mining industry in bioleaching. Biotechnology is also used to recycle, treat waste, clean up sites contaminated by industrial activities (bioremediation), and also to produce biological weapons.

A series of derived terms have been coined to identify several branches of biotechnology, for example:

  • Bioinformatics (also called 'gold biotechnology') is an interdisciplinary field that addresses biological problems using computational techniques, and makes the rapid organization as well as analysis of biological data possible. The field may also be referred to as computational biology, and can be defined as, 'conceptualizing biology in terms of molecules and then applying informatics techniques to understand and organize the information associated with these molecules, on a large scale.'[21] Bioinformatics plays a key role in various areas, such as functional genomics, structural genomics, and proteomics, and forms a key component in the biotechnology and pharmaceutical sector.[22]
  • Blue biotechnology is based on the exploitation of sea resources to create products and industrial applications.[23] This branch of biotechnology is the most used for the industries of refining and combustion principally on the production of bio-oils with photosynthetic micro-algae.[23][24]
  • Green biotechnology is biotechnology applied to agricultural processes. An example would be the selection and domestication of plants via micropropagation. Another example is the designing of transgenic plants to grow under specific environments in the presence (or absence) of chemicals. One hope is that green biotechnology might produce more environmentally friendly solutions than traditional industrial agriculture. An example of this is the engineering of a plant to express a pesticide, thereby ending the need of external application of pesticides. An example of this would be Bt corn. Whether or not green biotechnology products such as this are ultimately more environmentally friendly is a topic of considerable debate.[23] It is commonly considered as the next phase of green revolution, which can be seen as a platform to eradicate world hunger by using technologies which enable the production of more fertile and resistant, towards biotic and abiotic stress, plants and ensures application of environmentally friendly fertilizers and the use of biopesticides, it is mainly focused on the development of agriculture.[23] On the other hand, some of the uses of green biotechnology involve microorganisms to clean and reduce waste.[25][23]
  • Red biotechnology is the use of biotechnology in the medical and pharmaceutical industries, and health preservation.[23] This branch involves the production of vaccines and antibiotics, regenerative therapies, creation of artificial organs and new diagnostics of diseases.[23] As well as the development of hormones, stem cells, antibodies, siRNA and diagnostic tests.[23]
  • White biotechnology, also known as industrial biotechnology, is biotechnology applied to industrial processes. An example is the designing of an organism to produce a useful chemical. Another example is the using of enzymes as industrial catalysts to either produce valuable chemicals or destroy hazardous/polluting chemicals. White biotechnology tends to consume less in resources than traditional processes used to produce industrial goods.[26][27]
  • 'Yellow biotechnology' refers to the use of biotechnology in food production, for example in making wine, cheese, and beer by fermentation.[23] It has also been used to refer to biotechnology applied to insects. This includes biotechnology-based approaches for the control of harmful insects, the characterisation and utilisation of active ingredients or genes of insects for research, or application in agriculture and medicine and various other approaches.[28]
  • Gray biotechnology is dedicated to environmental applications, and focused on the maintenance of biodiversity and the remotion of pollutants.[23]
  • Brown biotechnology is related to the management of arid lands and deserts. One application is the creation of enhanced seeds that resist extreme environmental conditions of arid regions, which is related to the innovation, creation of agriculture techniques and management of resources.[23]
  • Violet biotechnology is related to law, ethical and philosophical issues around biotechnology.[23]
  • Dark biotechnology is the color associated with bioterrorism or biological weapons and biowarfare which uses microorganisms, and toxins to cause diseases and death in humans, livestock and crops.[29][23]

Medicine[edit]

In medicine, modern biotechnology has many applications in areas such as pharmaceutical drug discoveries and production, pharmacogenomics, and genetic testing (or genetic screening).

DNA microarray chip – some can do as many as a million blood tests at once

Pharmacogenomics (a combination of pharmacology and genomics) is the technology that analyses how genetic makeup affects an individual's response to drugs.[30] Researchers in the field investigate the influence of genetic variation on drug responses in patients by correlating gene expression or single-nucleotide polymorphisms with a drug's efficacy or toxicity.[31] The purpose of pharmacogenomics is to develop rational means to optimize drug therapy, with respect to the patients' genotype, to ensure maximum efficacy with minimal adverse effects.[32] Such approaches promise the advent of 'personalized medicine'; in which drugs and drug combinations are optimized for each individual's unique genetic makeup.[33][34]

Biotechnology Note Pdf
Computer-generated image of insulin hexamers highlighting the threefold symmetry, the zinc ions holding it together, and the histidine residues involved in zinc binding.

Biotechnology has contributed to the discovery and manufacturing of traditional small moleculepharmaceutical drugs as well as drugs that are the product of biotechnology – biopharmaceutics. Modern biotechnology can be used to manufacture existing medicines relatively easily and cheaply. The first genetically engineered products were medicines designed to treat human diseases. To cite one example, in 1978 Genentech developed synthetic humanized insulin by joining its gene with a plasmid vector inserted into the bacterium Escherichia coli. Insulin, widely used for the treatment of diabetes, was previously extracted from the pancreas of abattoir animals (cattle or pigs). The genetically engineered bacteria are able to produce large quantities of synthetic human insulin at relatively low cost.[35][36] Biotechnology has also enabled emerging therapeutics like gene therapy. The application of biotechnology to basic science (for example through the Human Genome Project) has also dramatically improved our understanding of biology and as our scientific knowledge of normal and disease biology has increased, our ability to develop new medicines to treat previously untreatable diseases has increased as well.[36]

Genetic testing allows the geneticdiagnosis of vulnerabilities to inherited diseases, and can also be used to determine a child's parentage (genetic mother and father) or in general a person's ancestry. In addition to studying chromosomes to the level of individual genes, genetic testing in a broader sense includes biochemical tests for the possible presence of genetic diseases, or mutant forms of genes associated with increased risk of developing genetic disorders. Genetic testing identifies changes in chromosomes, genes, or proteins.[37] Most of the time, testing is used to find changes that are associated with inherited disorders. The results of a genetic test can confirm or rule out a suspected genetic condition or help determine a person's chance of developing or passing on a genetic disorder. As of 2011 several hundred genetic tests were in use.[38][39] Since genetic testing may open up ethical or psychological problems, genetic testing is often accompanied by genetic counseling.

Agriculture[edit]

Genetically modified crops ('GM crops', or 'biotech crops') are plants used in agriculture, the DNA of which has been modified with genetic engineering techniques. In most cases, the main aim is to introduce a new trait that does not occur naturally in the species. Biotechnology firms can contribute to future food security by improving the nutrition and viability of urban agriculture. Furthermore, the protection of intellectual property rights encourages private sector investment in agrobiotechnology. For example, in Illinois FARM Illinois (Food and Agriculture RoadMap for Illinois) is an initiative to develop and coordinate farmers, industry, research institutions, government, and nonprofits in pursuit of food and agriculture innovation. In addition, the Illinois Biotechnology Industry Organization (iBIO) is a life sciences industry association with more than 500 life sciences companies, universities, academic institutions, service providers and others as members. The association describes its members as 'dedicated to making Illinois and the surrounding Midwest one of the world’s top life sciences centers.'[40]

Examples in food crops include resistance to certain pests,[41] diseases,[42] stressful environmental conditions,[43] resistance to chemical treatments (e.g. resistance to a herbicide[44]), reduction of spoilage,[45] or improving the nutrient profile of the crop.[46] Examples in non-food crops include production of pharmaceutical agents,[47]biofuels,[48] and other industrially useful goods,[49] as well as for bioremediation.[50][51]

Farmers have widely adopted GM technology. Between 1996 and 2011, the total surface area of land cultivated with GM crops had increased by a factor of 94, from 17,000 square kilometers (4,200,000 acres) to 1,600,000 km2 (395 million acres).[52] 10% of the world's crop lands were planted with GM crops in 2010.[52] As of 2011, 11 different transgenic crops were grown commercially on 395 million acres (160 million hectares) in 29 countries such as the US, Brazil, Argentina, India, Canada, China, Paraguay, Pakistan, South Africa, Uruguay, Bolivia, Australia, Philippines, Myanmar, Burkina Faso, Mexico and Spain.[52]

Genetically modified foods are foods produced from organisms that have had specific changes introduced into their DNA with the methods of genetic engineering. These techniques have allowed for the introduction of new crop traits as well as a far greater control over a food's genetic structure than previously afforded by methods such as selective breeding and mutation breeding.[53] Commercial sale of genetically modified foods began in 1994, when Calgene first marketed its Flavr Savr delayed ripening tomato.[54] To date most genetic modification of foods have primarily focused on cash crops in high demand by farmers such as soybean, corn, canola, and cotton seed oil. These have been engineered for resistance to pathogens and herbicides and better nutrient profiles. GM livestock have also been experimentally developed; in November 2013 none were available on the market,[55] but in 2015 the FDA approved the first GM salmon for commercial production and consumption.[56]

There is a scientific consensus[57][58][59][60][61][62][63] that currently available food derived from GM crops poses no greater risk to human health than conventional food,[64][65][66][67][68][69][70] but that each GM food must be tested on a case-by-case basis before introduction.[71][72][73] Nonetheless, members of the public are much less likely than scientists to perceive GM foods as safe.[74][75][76][77] The legal and regulatory status of GM foods varies by country, with some nations banning or restricting them, and others permitting them with widely differing degrees of regulation.[78][79][80][81]

GM crops also provide a number of ecological benefits, if not used in excess.[82] However, opponents have objected to GM crops per se on several grounds, including environmental concerns, whether food produced from GM crops is safe, whether GM crops are needed to address the world's food needs, and economic concerns raised by the fact these organisms are subject to intellectual property law.

Industrial[edit]

Industrial biotechnology (known mainly in Europe as white biotechnology) is the application of biotechnology for industrial purposes, including industrial fermentation. It includes the practice of using cells such as microorganisms, or components of cells like enzymes, to generate industrially useful products in sectors such as chemicals, food and feed, detergents, paper and pulp, textiles and biofuels.[83] In the current decades, significant progress has been done in creating genetically modified organisms (GMOs) that enhance the diversity of applications and economical viability of industrial biotechnology. By using renewable raw materials to produce a variety of chemicals and fuels, industrial biotechnology is actively advancing towards lowering greenhouse gas emissions and moving away from a petrochemical-based economy.[84]

Environmental[edit]

The environment can be affected by biotechnologies, both positively and adversely. Vallero and others have argued that the difference between beneficial biotechnology (e.g.bioremediation is to clean up an oil spill or hazard chemical leak) versus the adverse effects stemming from biotechnological enterprises (e.g. flow of genetic material from transgenic organisms into wild strains) can be seen as applications and implications, respectively.[85] Cleaning up environmental wastes is an example of an application of environmental biotechnology; whereas loss of biodiversity or loss of containment of a harmful microbe are examples of environmental implications of biotechnology.

Regulation[edit]

The regulation of genetic engineering concerns approaches taken by governments to assess and manage the risks associated with the use of genetic engineering technology, and the development and release of genetically modified organisms (GMO), including genetically modified crops and genetically modified fish. There are differences in the regulation of GMOs between countries, with some of the most marked differences occurring between the USA and Europe.[86] Regulation varies in a given country depending on the intended use of the products of the genetic engineering. For example, a crop not intended for food use is generally not reviewed by authorities responsible for food safety.[87] The European Union differentiates between approval for cultivation within the EU and approval for import and processing. While only a few GMOs have been approved for cultivation in the EU a number of GMOs have been approved for import and processing.[88] The cultivation of GMOs has triggered a debate about coexistence of GM and non GM crops. Depending on the coexistence regulations, incentives for cultivation of GM crops differ.[89]

Learning[edit]

In 1988, after prompting from the United States Congress, the National Institute of General Medical Sciences (National Institutes of Health) (NIGMS) instituted a funding mechanism for biotechnology training. Universities nationwide compete for these funds to establish Biotechnology Training Programs (BTPs). Each successful application is generally funded for five years then must be competitively renewed. Graduate students in turn compete for acceptance into a BTP; if accepted, then stipend, tuition and health insurance support is provided for two or three years during the course of their Ph.D. thesis work. Nineteen institutions offer NIGMS supported BTPs.[90] Biotechnology training is also offered at the undergraduate level and in community colleges.

See also[edit]

  • List of biotechnology companies

References and notes[edit]

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  89. ^Beckmann VC, Soregaroli J, Wesseler J (2011). 'Coexistence of genetically modified (GM) and non-modified (non GM) crops: Are the two main property rights regimes equivalent with respect to the coexistence value?'. In Carter C, Moschini G, Sheldon I (eds.). Genetically modified food and global welfare. Frontiers of Economics and Globalization Series. 10. Bingley, UK: Emerald Group Publishing. pp. 201–224.
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Further reading[edit]

  • Friedman Y (2008). Building Biotechnology: Starting, Managing, and Understanding Biotechnology Companies. Washington, DC: Logos Press. ISBN978-0-9734676-3-5.
  • Oliver RW (2000). The Coming Biotech Age. ISBN978-0-07-135020-4.
  • Powell WW, White DR, Koput KW, Owen-Smith J (2005). 'Network Dynamics and Field Evolution: The Growth of Interorganizational Collaboration in the Life Sciences'. American Journal of Sociology. 110 (4): 1132–1205. CiteSeerX10.1.1.319.1227. doi:10.1086/421508. Viviana Zelizer Best Paper in Economic Sociology Award (2005–2006), American Sociological Association.
  • Rasmussen N (2014). Gene Jockeys: Life Science and the rise of Biotech Enterprise. Baltimore, MD: Johns Hopkins University Press.
  • Zaid A, Hughes HG, Porceddu E, Nicholas F (2001). Glossary of Biotechnology for Food and Agriculture — A Revised and Augmented Edition of the Glossary of Biotechnology and Genetic Engineering. Available in English, French, Spanish, Chinese, Arabic, Russian, Polish, Serbian, Vietnamese and Kazakh. Rome: FAO. ISBN978-92-5-104683-8.
  • Caswell MF, Fuglie KO, Klotz CA (1998). 'Agricultural Biotechnology: An Economic Perspective'. Agricultural Economic Report. United States Department of Agriculture Economic Research Service.

External links[edit]

Basic Biotechnology Pdf

Wikibooks has a book on the topic of: Genes, Technology and Policy
At Wikiversity, you can learn more and teach others about Biotechnology at the Department of Biotechnology
Look up biotechnology in Wiktionary, the free dictionary.
  • Media related to Biotechnology at Wikimedia Commons
  • Foundation for Biotechnology Awareness and Education,
  • A report on Agricultural Biotechnology focusing on the impacts of 'Green' Biotechnology with a special emphasis on economic aspects. fao.org.
  • US Economic Benefits of Biotechnology to Business and Society NOAA Economics, economics.noaa.gov
  • Database of the Safety and Benefits of Biotechnology – a database of peer-reviewed scientific papers and the safety and benefits of biotechnology.
Retrieved from 'https://en.wikipedia.org/w/index.php?title=Biotechnology&oldid=920506153'

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CBSE Class 12 Biology Revision Notes Chapter 11 Biotechnology Principles and Processes

The techniques of using live organisms or enzymes from organisms to produce products and processes useful to humans. Many processes like in vitro fertilization leading to ‘test-tube’ baby, synthesizing gene and using it, developing a DNA vaccine or correcting a defective gene are also parts of Biotechnology.

The European Federation of Biotechnology (EFB) has given a definition of biotechnology that comprises both traditional and modern molecular biotechnology.The definition is as follow- “The integration of natural science and organisms, cells, parts thereof, and molecular analogous for products and services”.

Principles of Biotechnology

Modern biotechnology is based on two main principles-

• Genetic Engineering – Genetic Engineering is defined as the direct manipulation of genome (DNA and RNA) of an organism. It involves the transfer of new genes to improve the function or trait into host organisms and thus changes the phenotype of the host organism.

• Maintenance of sterile condition in chemical engineering process to enable growth of only desired microbes for manufacture of biotechnological products like antibiotics, vaccine, enzymes etc.

• Traditional hybridization used in plants and animal breeding leads to inclusion and multiplication of undesirable genes along with the desired traits. The technique of genetic engineering which include creation of recombinant DNA, use of gene cloning and gene transfer allow us to isolate and introduce only one or a set of desirable genes without introducing undesirable genes into the target organism.

• In a chromosome there is a specific DNA sequence called the origin of replication, which is responsible for initiating replication. Therefore, for the multiplication of any alien piece of DNA in an organism, it needs to be a part of a chromosome which has a specific sequence known as ‘origin of replication’. Thus, an alien DNA is linked with the origin of replication, so that, this alien piece of DNA can replicate and multiply itself in the host organism. This is known as Cloning or making multiple identical copies of any template DNA.

• The construction of the first recombinant DNA emerged from the possibility of linking a gene encoding antibiotic resistance with a native Plasmid of Salmonella typhimurium.

  • Stanley Cohen and Herbert Boyer in 1972 isolated the antibiotic resistance gene by cutting out a piece of DNA from a plasmid (autonomously replicating circular extra-chromosomal DNA) of Salmonella typhimurium. The cutting of DNA at specific locations became possible with the discovery of the so-called ‘molecular scissors’– restriction enzymes.

• The cut piece of DNA was then linked with the plasmid DNA. These plasmid DNA act as vectors to transfer the piece of DNA attached to it.A plasmid can be used as vector to deliver an alien piece of DNA into the host organism.

• The linking of antibiotic resistance gene with the plasmid vector become possible with the enzyme ligase, which acts on cut DNA molecules and joins their ends. This makes a new combination of autonomously replicating DNA created in vitro and known as recombinant DNA.

• When this DNA is transferred into E.coli, it could replicate using the new host DNA polymerase enzyme and make multiple copies. The ability to multiply copies of antibiotic resistance gene in E.coli was called cloning of antibiotic resistance gene in E.coli.

“Recombinant DNA technology” or also called “Genetic Engineering” deals about, the production of new combinations of genetic material (artificially) in the laboratory. These “recombinant DNA” (rDNA) molecules are then introduced into host cells, where they can be propagated and multiplied.

Steps of Fecombinant DNA Technology –

I. Identification of DNA with desirable genes.

II. Introduction of the identified DNA into the host.

III. Maintenance of introduced DNA in the host and transfer of the DNA to its progeny.

Tools of Recombinant DNA Technology includes

• Restriction Enzymes

• Polymerase enzymes

• Ligases

• Vectors

• Host organisms

Restriction Enzymes (Molecular Scissors):

Restriction enzymes belong to a larger class of enzymes called Nucleases. There are of two kinds; Exonucleases and Endonucleases. Exonucleases remove nucleotides from the ends of the DNA whereas, endonucleases make cuts at specific position within the DNA.

Example, the first restriction endonuclease – Hind II, always cut DNA molecules at a particular point by recognizing a specific sequence of six base pairs. This specific base sequence is known as the Recognition Sequence for Hind II.

Biotechnology Powerpoint For High School

• Each restriction endonuclease recognises a specific palindromic nucleotide sequence in the DNA. Palindromes are group of letters that form the same words when read both forward and backward for example “MALYALAM”.

5′ —— GAATTC —— 3′

3′ —— CTTAAG —— 5′

The palindrome in DNA is a sequence of base pairs that reads same on two stands when orientation of reading is kept the same.

• Restriction enzymes cut the strand of DNA a little away from the centre of the palindrome site between the same two bases on the opposite strands having sticky strand. The stickiness of the strands facilities the action of the enzyme DNA ligase.

• Restriction endonucleases are used in genetic engineering to form recombinant molecules of DNA which are composed of DNA from different sources or genome.

• When cut the same restriction enzyme the resultant DNA fragments have the same kind of Sticky-ends and can be joined together using DNA ligases.

Diagrammatic representation of Recombinant DNA technology

Separation and isolation of DNA fragments

The fragment of DNA obtained by cutting DNA using restriction enzyme is separated by technique called gel electrophoresis. Negatively charged DNA fragments can be separated by forcing them to move towards the anode under an electric field through medium. DNA fragments separate according to their size through sieving effect provided by agarose gel.

• The separated DNA fragment can be visualized after staining the DNA with ethodium bromide followed by exposure to UV light. Separated bands of DNA are separated from agarose gel and extracted from gel, called elution. The DNA fragment purified this way is used for recombination.

Cloning Vector

Plasmids and Bacteriophages is commonly used vector for cloning. They have ability to replicate within bacterial cells independent of the control of chromosomal DNA. Bacteriophages because of their high number per cell, have very high copy numbers of their genome within the bacterial cells.

Following features are required to facilitate cloning into a vector-

a. Origin of replication (ori) – the sequence from where replication starts and any piece of DNA when linked to this sequence can be made to replicate within the host cells.This sequence is responsible for controlling the copy number of the linked DNA.

b. Selectable marker-help in the identifying and eliminating non transformants and selectively permitting the growth of the transformants. Transformation is a procedure through which a piece of DNA is introduced in a host bacterium. Generally,the genes encoding resistance to antibiotics such as ampicillin, chloramphenicol, tetracycline or kanamycin, etc., are considered useful selectable markers for E. coli.

c. Cloning sites– to link the foreign DNA, the vector need to have single recognition sites for the commonly used restriction enzymes as presence of more than one recognition sites within the vector will generate several fragments, which will complicate the gene cloning. The ligation of foreign DNA is carried out at a restriction site present in one of the two antibiotic resistance genes.

E. coli cloning vector pBR322 showing restriction sites (Hind III, EcoR I, BamH I, Sal I, Pvu II, Pst I, Cla I), ori and antibiotic resistance genes (ampR and tetR ). rop codes for the proteins involved in the replication of the plasmid.

Insertional inactivation:
The most efficient method of screening for the presence of recombinant plasmids is based on the principle that the cloned DNA fragment disrupts the coding sequence of a gene. This is termed as Insertional Inactiviation.

For example, the powerful method of screening for the presence of recombinant plasmids is referred to as Blue-White selection. This method is based upon the insertional inactivation of the lac Z gene present on the vector. The lac Z gene encodes the enzyme beta-galactosidase, which can cleave a chromogenic substrate into a blue coloured product. If this lac Z gene is inactivated by insertion of a target DNA fragment into it, the development of the blue colour will be prevented and it gives white coloured colonies. By this way, we can differentiate recombinant (white colour) and non-recombinant (blue colour) colonies.

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d. Vectors for cloning genes in plants and animals– Agrobacterium tumefactions (pathogen of dicot plant) is able to deliver a piece of DNA known as ‘T-DNA” to transform normal plant cells into a tumor and direct these tumor cells to produce the chemicals required by the pathogen. Retroviruses in animals have the ability to transform normal cells into cancerous cells. The tumor inducing (Ti) plasmid of Agrobacterium tumefaciens has been modified into cloning vector having no more pathogenic to plant. Similarly retrovirus have been modified into cloning vector for animals.

Competent host (For Transformation with Recombinant DNA)

1) Simple chemical treatment with divalent calcium ions increases the efficiency of host cells (through cell wall pores) to take up the rDNA plasmids.
2) rDNA can also be transformed into host cell by incubating both on ice, followed by placing them briefly at 42oC (Heat Shock), and then putting them back on ice. This enables the bacteria to take up the recombinant DNA.
3) In Microinjection method, rDNA is directly injected into the nucleus of cells by using a glass micropipette.
4) Biolistics / Gene gun method, it has been developed to introduce rDNA into mainly plant cells by using a Gene / Particle gun. In this method, microscopic particles of gold / tungsten are coated with the DNA of interest and bombarded onto cells.
5) The last method uses “Disarmed Pathogen” Vectors (Agrobacterium tumefaciens), which when allowed to infect the cell, transfer the recombinant DNA into the host.

Processes of Recombinant DNA Technology

Recombinant DNA technology involves several steps in specific sequence-

a. Isolation of DNA

b. Fragmentation of DNA by restriction endonucleases

c. Isolation of a desired DNA fragment

d. Ligation of the DNA fragment into vector

e. Transforming the recombinant DNA into the host

f. Culturing the host cells in a medium at large scale

g. Extraction of the desired product.

•Isolation of Genetic material:Genetic material is isolated from other macromolecules by using enzymes such as lysozyme (bacteria), cellulase (plant cells), chitinase (fungus). DNA that separate out can be removed by spooling. The RNA can be removed by treatment with ribonuclease whereas proteins can be removed by treatment with protease.

• Cutting of DNA at specific location is performed by using restriction enzyme and Agarose gel electrophoresis to check the progression of a restriction enzyme digestion. After cutting sources of DNA as well as vector DNA with a specific restriction enzyme to cut out ‘gene of interest’ from the source DNA.

• Amplification of Gene of Interest using PCR( Polymerase Chain Reaction) to get multiple copies of the DNA or gene of interest in vitro by using set of primers and enzyme DNA polymerase.

Polymerase chain reaction (PCR) : Each cycle has three steps: (A) Denaturation; (B) Primer annealing; and (C) Extension of primers

This repeated amplification is done by the use of a thermostable DNA polymerase (isolated from a bacterium, Thermus aquaticus), which remain active during the high temperature induced denaturation of double stranded DNA.

• Insertion of Recombinant DNA into the Host Cell/Organism includes making the recipient cells competent to receive, take up DNA present in its surrounding etc. The recombinant DNA bearing gene for resistance to an antibiotic is transferred into E.coli cells, the host cell become transformed into ampicillin-resistance cells.

• Obtaining the foreign gene product – the foreign DNA multiplies in plant or animal cell to produce desirable protein. Expression of foreign genes in host cells involve, optimized condition to obtain recombinant protein. The recombinant cell is multiplied in a continuous culture system in which used medium is drained out from one side while fresh medium is added from the other to maintain the cells in their physiological active phase. A bioreactor provides the optimal conditions for achieving the desired product by providing optimum growth conditions (temperature, pH, substrate, salts, vitamins, oxygen).

• Downstream Processing involves processes that make the product obtain ready for marketing. This process includes separation and purification called as downstream processing. Suitable preservatives are added to it and send for clinical trial in case of drugs before releasing to market for public use

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