Department of Genetic Engineering and Biotechnology

Shahjalal University of Science and Technology

Sylhet -3114, Bangladesh

Course No. GEB 121

Course Title: Introduction To Genetic Engineering and Biotechnology

Submitted to :
Prof. Dr. Md Abdullah Al Mamun
Department of Genetic Engineering and Biotechnology
Shahjalal University of Science and Technology
Sylhet -3114, Bangladesh.

Submitted by :
Name : Sharmista Deb
Reg. No. : 2019451001
Session : 2019-2020
Department of Genetic Engineering and Biotechnology
 Shahjalal University of Science and Technology
Sylhet -3114, Bangladesh.

Genetic Engineering & Biotechnology

Genetic engineering, also called genetic modification, is the direct manipulation of an organisms
genome using biotechnology. It is a set of technologies used to change the genetic makeup of
cells, including the transfer of genes within and across species boundaries to produce improved
or novel organisms. New DNA is obtained by either isolating or copying the genetic material of
interest using molecular cloning methods or by artificially synthesizing the DNA. A construct is
usually created and used to insert this DNA into the host organism. As well as inserting genes,
the process can also be used to remove, or "knock out", genes. The new DNA can be inserted
randomly, or targeted to a specific part of the genome.

Biotechnology
Biotechnology is a broad area of biology, involving the use of living systems and organisms to
develop or make products. Depending on the tools and applications, it often overlaps with related
scientific fields. 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. The term "Biotechnology" was first
used by "Karl Ereky" in 1919, meaning the production of products from raw materials with the aid of
living organisms.

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.[1] 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.[2] Biotechnology is based on the basic biological
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).[3][4][5] The utilization of biological processes, organisms or systems to
produce products that are anticipated to improve human lives is termed biotechnology. [6]
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. [7] 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
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.[8] 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,e 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.[9]
For thousands of years, humans have used selective breeding to improve the 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. [10]
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.[11]
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.[10]
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
modified microorganism could be patented in the case of Diamond v. Chakrabarty.[12] 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.
The MOSFET (metal-oxide-semiconductor field-effect transistor) was invented by Mohamed M.
Atalla and Dawon Kahng in 1959.[13] Two years later, Leland C. Clark and Champ Lyons invented the
first biosensor in 1962.[14][15] Biosensor MOSFETs 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][18] It
is a special type of MOSFET,[16] where the metal gate is replaced by an ion-
sensitive membrane, electrolyte solution and reference electrode.[19] 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.[19]
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.[19]
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.[20]
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. [21]

Applications Of Biotechnology

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." [22] 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.[23]
 Blue biotechnology is based on the exploitation of sea resources to create products and
industrial applications.[24] 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.[24][25]
 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. [24] 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.[24] On the other hand, some of the uses of green biotechnology
involve microorganisms to clean and reduce waste.[26][24]
 Red biotechnology is the use of biotechnology in the medical
and pharmaceutical industries, and health preservation.[24] This branch involves the
production of vaccines and antibiotics, regenerative therapies, creation of artificial
organs and new diagnostics of diseases.[24] As well as the development
of hormones, stem cells, antibodies, siRNA and diagnostic tests.[24]
 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.[27][28]
 "Yellow biotechnology" refers to the use of biotechnology in food production, for example
in making wine, cheese, and beer by fermentation.[24] 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.[29]
 Gray biotechnology is dedicated to environmental applications, and focused on the
maintenance of biodiversity and the remotion of pollutants.[24]
 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.[24]
 Violet biotechnology is related to law, ethical and philosophical issues around
biotechnology.[24]
  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.[30][24]
Medicine
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. [31] 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.[32] 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.[33] Such approaches
promise the advent of "personalized medicine"; in which drugs and drug combinations are optimized
for each individual's unique genetic makeup. [34][35]

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

molecule pharmaceutical 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.[36][37] 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. [37]
Genetic testing allows the genetic diagnosis 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.[38] 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.[39][40] Since genetic testing
may open up ethical or psychological problems, genetic testing is often accompanied by genetic
counseling.

Agriculture
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."[41]
Examples in food crops include resistance to certain pests,[42] diseases,[43] stressful environmental
conditions,[44] resistance to chemical treatments (e.g. resistance to a herbicide[45]), reduction of
spoilage,[46] or improving the nutrient profile of the crop.[47] Examples in non-food crops include
production of pharmaceutical agents,[48] biofuels,[49] and other industrially useful goods,[50] as well as
for bioremediation.[51][52]
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).[53] 10% of the world's crop lands were planted
with GM crops in 2010.[53] 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.[53]
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.
[54]
 Commercial sale of genetically modified foods began in 1994, when Calgene first marketed
its Flavr Savr delayed ripening tomato.[55] 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,[56] but in 2015 the FDA approved the first GM salmon for commercial
production and consumption.[57]
There is a scientific consensus[58][59][60][61] that currently available food derived from GM crops poses no
greater risk to human health than conventional food, [62][63][64][65][66] but that each GM food needs to be
tested on a case-by-case basis before introduction. [67][68][69] Nonetheless, members of the public are
much less likely than scientists to perceive GM foods as safe.[70][71][72][73] 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. [74][75][76][77]
GM crops also provide a number of ecological benefits, if not used in excess. [78] 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
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.[79] 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. [80]

Environmental
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. [81] 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.

Biotechnology And Developing Countries

It is now widely recognised by industrialised and developing countries alike that
technology and the successful adaptation of new technologies is crucial for economic
growth and sustainable development. The wide range of techniques presented by
biotechnology provide an important opportunity for developing countries to adopt
technologies which can be tailored to their own environmental and societal needs in a
sustainable manner. The successful adaptation of a science-intensive technology such
as biotechnology, however, involves a number of layers of management, including the
development of the basic sciences, institutional capabilities and the establishment of a
macroenvironment which is conducive to technological change and production.

The basic task of this paper is to assess national technological capabilities for
biotechnology management in developing countries. The paper is concerned with
what constitutes a nation's capabilities for managing the development and application
of biotechnology as well as guiding its evolution in local socioeconomic systems. We
examine national policies to develop biotechnology in a number of countries in south
and south-east Asia and Africa and compare their experiences in an attempt to
determine what their particular strengths and weaknesses are. Due to the different
levels of capability found in these parts of the world as well as different industrial
structures across sectors, the tendency has been to concentrate on developing different
areas of specialisation. Thus, technological change has been incremental rather than
radical in both these areas. However, Asia and especially the newly industrialising
countries of Southeast Asia have a tendency to race ahead of many other countries, as
would be expected, because of their previous accumulation of technological
capabilities. Nevertheless, the fact that even these countries have had differing
degrees of success in commercialising biotechnology in different sectors, shows that
possessing scientific capability to adapt technology is not sufficient. Instead,
capabilities at different levels, to innovate and also to efficiently manage the
technology must be developed in order for developing countries to register success in
harnessing biotechnology for their own needs and priorities.

Commercialization Of Biotechnology In A Developing Country

In general, commercialization of agricultural biotechnology is far behind the

application of biotechnology in human health and medicine. In the developed
countries, especially in Japan, the private sector plays a major role in the generation,
development and commercialization of biotechnology products and techniques.
Because of commercial considerations, concentration has been mainly on diagnostics,
vaccines, pharmaceuticals and other health-related products. Moreover, because of the
eminent involvement of the private sector, marketability of the products and potential
return on investment are crucial factors in deciding which products are to be
developed and commercialized.
In Japan, the MAFF is subsidizing fundamental technical development projects in the
private sector and providing funds for investment in such projects. It is also offering
financial assistance to prefectural governments so as to encourage their R&D
activities and utilization of the outcome of such activities. The Ministry takes steps to
subsidize the research projects of private technical research associations and other
similar study programmes. These subsidies are offered mainly through the Ministry's
project for the development of biotechnology and other high technology for the food
industry. In fiscal 1989, MAFF began such financial assistance programmes as the
"Transformation of plant cells through the introduction of small organelles into the
cells" and "Genetically manipulated vaccines against animal protozoa diseases".

In the area of biotechnological R&D activities in the food industry, the Ministry
started the project "Development of bioreactor systems for the food industry" in 1984.
This project has achieved a number of remarkable results regarding the conversion of
saccharides, fat and other substances. Another project, the "Development of
technology for improvement of enzyme functions for the food industry" aims to
improve the functions of enzymes for food production utilizing recombinant DNA
technology. In fiscal 1989, the Ministry began a new programme, "Food production
by large-scale, high-density fermentation under ultra-high pressure conditions" with a
view to increasing the efficiency of the food production process under ultra-high
pressures and to finding food materials with a higher value added.

Two more projects were launched in 1989: the "Utilization of extracts from trees
(green sprouts project)" and "Advanced and multipurpose utilization of unused marine
resources (marine-frontier project)". The objective of these projects is to develop the
technique for extracting useful substances from trees and marine organisms and
utilizing them for foods, perfumes and medicines, thereby helping further growth of
forestry and fishery industries.

The Bio-Oriented Technology Research Advancement Institution (BRAIN) was

founded in October 1986 by the joint investment of the government and the private
sector. In an effort to encourage the R&D activities of the industries related to
agriculture, forestry, fishery and food production, this institution is investing in joint
technical development corporations and is providing no interest-bearing loans to the
technical research projects of businesses on certain conditions.

During three years from fiscal 1986 to 1988, BRAIN invested in 14 study projects and
offered loans to 70 projects. In fiscal 1989, the Institution invested in, among others,
the Institute of Aquacultural Technology of Cold Water Fishes, Wakayama Agro-
biological Research Centre and the Japan Turf Grass Co. The first is engaged in the
development of aquaculture systems of cold water high-class fishes, the second in
R&D activities for rationalized process of tangerine juice production and the third in
the breeding of better turfs.

In the area of biotechnology-related R&D in the private sector, financing systems

through the Japan Development Bank, the Agriculture, Forestry and Fishery Finance
Corporation and other similar government organizations were reinforced to turn the
results of these R&D activities to practical use. It is hoped that as biotechnology is
increasingly introduced to industries, these financial measures will be utilized more in
the coming years.

Prefectures are becoming more and more interested in R&D activities on

biotechnology and the practical use of the outcome of such activities. This is evident
from the fact that almost all prefectures throughout Japan have a council on this
technology composed of scholars and other people of expertise.

MAFF is offering to prefectures a variety of subsidies in order to diffuse the mass

production technology of virus-free seedlings of vegetables, ornamental plants and
fruit trees and the transplanting technology of the embryos of beef cattle. In fiscal
1989, the Ministry started the "Project for establishing embryo supply centres" to
ensure a stable supply of embryos of beef cattle. This project is improving the raising
and controlling facilities of cattle from which such embryos are taken.

Since 1986, MAFF has also been implementing the "Project for assisting local R&D
activities in biotechnology". This project offers financial assistance to the R&D
activities of prefectural research institutes concerning the breeding and utilization
technology of biological resources. Other biotechnology-related activities of the
Ministry include regional biotechnology meetings held in seven locations across the
country with the object of exchanging information on R&D. The Ministry's new
initiative in fiscal 1989 included surveys and studies for examining the direction of
the practical use of biotechnology in each region. The Tsukuba Bioscience Hall
created in 1989 provides a forum for collaboration among trainees from industries,
universities and the government and international exchange programmes.
Furthermore, to foster links among genetic resources and biotechnology, the Ministry
has a strong national genebank and germplasm conservation programme.

Of the developed countries in the region, the involvement of the private sector in
biotechnology, especially plant biotechnology, is rather limited in New Zealand. Very
little research on plant biotechnology is being carried out by private companies. A
shortage of venture capital, low taxation incentives and tough competition from large
overseas firms make such ventures unattractive. Several small companies are engaged
in either product formulation, or manufacture of biological control agents. Larger
international companies (e.g. Monsanto, MPI Koln, PGS Ghent, ICI) and overseas
research institutions (UCLA, California; John Innes Institute, United Kingdom;
Christian Albrecht University, Germany) have provided personnel, facilities and some
supportive funding on collaborative projects. Producer Boards (e.g. New Zealand
Kiwifruit Marketing Board) also support research through joint funding of
government programmes.

Many New Zealand research programmes on plant biotechnology are not mature
enough to have reached the stage of successful commercialization. In addition, some
programmes are commercially sensitive, particularly where research is collaborative
with private companies. Examples of plant biotechnology research that have been, or
are in the process of being commercialized in New Zealand include:

 Establishment of a commercial tissue culture laboratory for clonal Forestry

with Radiata Pine (FRI).
 Use of immuno-molecular techniques for developing test assay kits and for
analysis of gene expression (DSIR, Fruit and Trees).
 Plant variety rights have been granted for four new plant varieties.

Future commercialization is likely to be through the release of improved crop

varieties, development of new cultivars and the licensing of technologies that have
been developed.

In the developing countries, in vitro culture for micropropagation of elite and disease-
free materials, production and distribution of efficient nitrogen-fixing microbes,
diagnostic kits and monoclonal antibody-based vaccines are the main areas that have
been commercialized to varying degrees in different countries.

In developing countries, the role of the private sector in modern biotechnology has
been rather limited. This is attributed partly to (i) vague government policies
regarding the private sector; (ii) unclear policies or no policies/views on patents and
intellectual property rights; (iii) poor links between public and private sectors; (iv)
low purchasing power for new products that are usually highly priced and are out of
reach of the majority of resource-poor farmers and low-income consumers; and (v)
inadequate local expertise and infrastructure and R&D support to new biotechnology.

In India, biotechnology industries have recently been growing in the private sector.
Hindustan Lever Ltd, an Indian subsidiary of the multinational Unilever, the Tata
Energy Research Institute (TERI) and Southern Petrochemical Industries are some of
the leading biotechnology companies in the country. These have considerable in-
house R&D bases active in the application of new technology to agribusiness, such as
enzyme technology for high-value chemicals and quality edible oil by using
genetically modified bacteria; protoplast fusion in yeast for efficient fermentation and
production of edible oil from biomass; tissue culture of plantation and ornamental
crops; biological nitrogen fixation; organic components for higher rate of
photosynthesis; big-insecticides; plant growth promoters; biomass processing for
animal feeds; and hybrid seeds. Some small and medium-size companies are also
initiating biotechnology activities but, with less R&D capacity, are more wary of what
they consider to be a higher-risk technology.

The private sector in India is particularly active in tissue culture products. There are
about 50 private sector companies such as Indo-American Hybrid Seeds, A.V.
Thomas and Co., Unicorn Biotek, the three companies mentioned above and others
that are comprehensively involved in commercial in vitro production of planting
materials of several horticultural and plantation crops such as cardamom, coffee, tea,
oilpalm, orchids, strawberry, roses, Spatiphyllum, banana, lily, Gerbera, etc. both for
domestic and export markets. These companies have established effective links with
the public sector.

The recent liberalization of trade, including that of certified seed, coupled with the
greater priority given to science and technology and the promotion of private sector
and public-private sector links, should encourage increased participation of the
industry in agricultural biotechnology R&D. Financial incentives are available for
supporting the growth of indigenous biotech industries.

Recombinant DNA Technology

Recombinant DNA Technology is defined by the Encyclopedia
Britannica as “the joining together of DNA molecules from different
organisms and inserting it into a host organism to produce new
genetic combinations that are of value to science, medicine,
agriculture and industry.”

Tools of Recombinant DNA technology

Inserting the desired gene into the genome of the host is not as easy as it sounds. It involves the
selection of the desired gene for administration into the host followed by a selection of the
perfect vector with which the gene has to be integrated and recombinant DNA formed. This
recombinant DNA then has to be introduced into the host. And at last, it has to be maintained in
the host and carried forward to the offspring.  Recombinant DNA technology can be complete
and achieved with the help of some elemental tools. The different tools used for the purpose are
discussed below:
Restriction Enzymes
 The restriction enzymes – help to cut, the polymerases- help to synthesize and the ligases- help
to bind.
The restriction enzymes used in recombinant DNA technology play a major role in determining
the location at which the desired gene is inserted into the vector genome. They are two types,
namely endonucleases and exonucleases. The endonucleases cut within the DNA strand whereas
the exonucleases cut the nucleotides from the ends of the DNA strands. The restriction
endonucleases are sequence-specific which is usually palindrome sequences and cut the DNA at
specific points. They scrutinize the length of DNA and make the cut at the specific site called the
restriction site. This gives rise to sticky ends in the sequence. The desired genes and the vectors
are cut by the same restriction enzymes to obtain the complementary sticky notes, thus making
the work of the ligases easy to bind the desired gene to the vector.

Vectors
The vectors help in carrying and integrating the desired gene. These form a very important part
of the tools of recombinant DNA technology as they are the ultimate vehicles that carry forward
the desired gene into the host organism. Plasmids and bacteriophages are the most common
vectors in recombinant DNA technology.
Host Organism
Host organism is the organism into which the recombinant DNA is introduced. The host is the
ultimate tool of recombinant DNA technology which takes in the vector engineered with the
desired DNA with the help of the enzymes. There are a number of ways in which this
recombinant DNA’s are inserted into the host, namely – microinjection, biolistics or gene gun,
alternate cooling and heating, use of calcium ions, etc.

Gin modification

Fred Couch's customers at Rabbit Ridge Gin in Lepanto, Ark., don't fully understand how the
powered paddle roll technology in the gin works, or that it increases capacity for the facility. All
they know is that they've seen an improvement in turnout and staple length since the technology
was installed three years ago.

The paddle roll device was invented by USDA/ARS engineer Weldon Laird at the Lubbock,
Texas, ARS facility, through research funded by Cotton Incorporated. It is a spinoff of Laird's
work on EasiFlo cottonseed — cottonseed which is treated with a starch and water coating after
ginning to make it easier to handle for dairies and feedlots.

Laird was researching ways to remove excess fiber, or tags, from the seed. USDA scientists
found that this extra fiber coming off the seed during the coating process resulted in clumping of
tangled seeds.

Laird set out the remove the tags by running ginned seed through a second gin with a powered
paddle roll and seed finger modifications. It worked so well that Laird started looking for a way
to retrofit the technology on existing gin stands — in effect to remove the excess fiber during the
original ginning process.

The successful venture now is marketed as the Power Roll gin stand and is being distributed
through PRT Marketing. The president of the company is Laird's son, Russell Laird.
The technology does not require the installation of a new gin stand. Instead the existing gin front,
or breast, is replaced with a new one that contains a larger seed roll area, a paddle roll to actively
turn the seed roll and a seed finger roll.

Couch believes that the increase in capacity at Rabbit Ridge is due to the Power Roll's addition
of a 30-horsepower motor to turn the seed roll. The gin has three 1973 Lummus 700 feeders and
three, early-1970s model Continental 141 gin stands, which do not have a seed roll turning
device.

“The ginning takes place by the saw causing the seed roll to begin to turn. So if I have 100
horsepower turning the saw, the 100 horses are also turning the seed roll.”

Adding the paddle roll technology allows all of the 100-horsepower motor to be used for turning
the saw. “The more power you can put on the saw, the more seedcotton you're going to put
through it. Our capacity increased a lot.”

In fact, capacity increased from 21.4 bales per hour in 2002 to 23.5 bales per hour in 2003, the
year the technology was added. That increased to 25.11 bales per hour the following year with
the addition of a higher-capacity press and moisture restoration. In 2005, capacity increased to
27.14 bales per hour with the installation of an improved gin front.

At Servico Gin, in Courtland, Ala., capacity increased from 26.5 bales per hour prior to
installation of the power roll to 29.2 bales per hour after installation, even though the gin stands
there do have a seed roll turning device.

Three years ago, the technology was installed on Murray 142 gin stands at Midnight Gin,
Midnight, Miss. Ginner Robert Royal says, “There were some nagging engineering problems,
but they were expected. But through it all, the seeds have been just buck-naked. That benefit
goes directly into the farmers' pockets.”
Initially, Royal wanted to increase capacity without losing turnout, but that wasn't accomplished
to his satisfaction until the end of the 2005 ginning season. “Weldon (Laird) and an engineer
made some additional modifications and all of a sudden, it started ginning cotton like we had
hoped it would, and turnout was still way up there. But it hasn't been through the paces of a full
season yet.”

Royal noted that fiber quality improvements due to the technology “were nominal, but there
wasn't any deterioration of quality.”

For Couch, the increase in capacity “has offset the labor costs and the increase in power and gas
to dry. Just being able to gin another one or two bales per hour has allowed us to keep our per-
bale costs stable.”

In addition, most of the gin's cotton producers — around 20 — “don't see nearly the deducts that
they did prior to the technology,” Couch said.

According to tests conducted by USDA, PRT Marketing and the gin, turnout in the three years
prior to installation of the technology averaged 35.7 percent. In the three years since the
technology was installed, turnout has averaged 38.2 percent.

“Normally, there will be a tail left on the seed during the ginning process,” Couch said. “That tail
essentially is excess lint. I was running somewhere around 12 percent residual lint left on the
seed prior to the Power Roll. With the technology, I got that under 10 percent. In cleaning the
seed better, that increased the return to the producer.”

The technology “won't take a 34 staple to a 35 staple,” Couch said. “But there will be more 35
staple length than what would come out of the conventional 141 Continental gin stand. I'm
seeing a significant improvement in turnout. I also reduced my short fiber content from 10
percent to 7.5 percent.”
Couch has an idea of how the technology works because he sees many of the same benefits from
slowing gin speed down considerably. “When you're ginning at a high rate, you're create a seed
roll that is really packed. Sometimes it's packed so tightly that the saw cannot pull the seed to the
ginning point. With the paddle roll technology keeping the flow of seedcotton broken up, every
time the lint touches the saw, it's able to take it to the gin point.”

Greg Holt, USDA/ARS agricultural engineer in Lubbock, says some research on some of the
Power Roll models has been mixed.”

On gin stand models where results were favorable, “studies showed that it did increase ginning
rate and turnout. There were some fiber properties that appeared to be better with the technology.

“Like any new technology coming in, there were some rough spots. The wrinkles are being
ironed out and the technology is turning the corner.”

Models that were having problems appear to have been fixed by a new design in 2005, according
to Holt. “But we haven't collected enough data to definitely say that we found the problem.”

Holt said the ginning laboratory in Lubbock is expecting to collect that data with the coming
ginning season.

“One of the potential benefits of the technology is the idea that it can control the speed of various
components in the gin stand,” Holt added. In the past, everything ran off one motor, which may
not have been the best for quality.

This gin stand could allow the ginner to change the speed of the individual components to
accommodate different cotton varieties. Some of the research indicates that on a varietal basis,
the setups probably need to be different.
Methods of Gene Transfer: 6 Methods
The six methods are: (1) Transformation (2) Conjugation
(3) Electroporation (4) Liposome-Mediated Gene Transfer
(5) Transduction and (6) Direct Transfer of DNA.
Method # 1. Transformation:
Transformation is the method of introducing foreign DNA into
bacterial cells (e.g. E.coli). The uptake of plasmid DNA by E.coli is
carried out in ice-cold CaCl2 (0-5°C), and a subsequent heat shock (37-
45°C for about 90 sec). By this technique, the transformation
frequency, which refers to the fraction of cell population that can be
transferred, is reasonably good e.g. approximately one cell for 1000
(10-3) cells.
Transformation efficiency:
It refers to the number of trans-formants per microgram of added
DNA. For E.coli, transformation by plasmid, the transformation
efficiency is about 107 to 108 cells per microgram of intact plasmid
DNA. The bacterial cells that can take up DNA are considered as
competent. The competence can be enhanced by altering growth
conditions.
The mechanism of the transformation process is not fully understood.
It is believed that the CaCI2 affects the cell wall, breaks at localized
regions, and is also responsible for binding of DNA to cell surface. A
brief heat shock (i.e. the sudden increase in temperature from 5°C to
40°C) stimulates DNA uptake. In general, large-sized DNAs are less
efficient in transforming.
Other chemical methods for transformation:
Calcium phosphate (in place of CaCI2) is preferred for the transfer of
DNA into cultured cells. Sometimes, calcium phosphate may result in
precipitate and toxicity to the cells. Some workers use diethyl amino
ethyl dextran (DEAE -dextran) for DNA transfer.
Method # 2. Conjugation:
Conjugation is a natural microbial recombination process. During
conjugation, two live bacteria (a donor and a recipient) come together,
join by cytoplasmic bridges and transfer single-stranded DNA (from
donor to recipient). Inside the recipient cell, the new DNA may
integrate with the chromosome (rather rare) or may remain free (as is
the case with plasmids).

Conjugation can occur among the cells from different genera of

bacteria (e.g Salmonella and Shigella cells). This is in contrast to
transformation which takes place among the cells of a bacterial genus.
Thus by conjugation, transfer of genes from two different and
unrelated bacteria is possible.

The natural phenomenon of conjugation is exploited for gene transfer.

This is achieved by transferring plasmid-insert DNA from one cell to
another. In general, the plasmids lack conjugative functions and
therefore, they are not as such capable of transferring DNA to the
recipient cells. However, some plasmids with conjugative properties
can be prepared and used.

Method # 3. Electroporation:
Electroporation is based on the principle that high voltage electric
pulses can induce cell plasma membranes to fuse. Thus,
electroporation is a technique involving electric field-mediated
membrane permeabilization. Electric shocks can also induce cellular
uptake of exogenous DNA (believed to be via the pores formed by
electric pulses) from the suspending solution.

Electroporation is a simple and rapid technique for introducing genes

into the cells from various organisms (microorganisms, plants and
animals).

The basic technique of electroporation for transferring genes into

mammalian cells is depicted in Fig. 6.11. The cells are placed in a
solution containing DNA and subjected to electrical shocks to cause
holes in the membranes. The foreign DNA fragments enter through
the holes into the cytoplasm and then to nucleus.
Electroporation is an effective way to transform E.coli cells containing
plasmids with insert DNAs longer than 100 kb. The transformation
efficiency is around 109 transformants per microgram of DNA for small
plasmids (about 3kb) and about 106 for large plasmids (about 130 kb).
Method # 4. Liposome-Mediated Gene Transfer:
Liposomes are circular lipid molecules, which have an aqueous
interior that can carry nucleic acids. Several techniques have been
developed to encapsulate DNA in liposomes. The liposome- mediated
gene transfer, referred to as lipofection, is depicted in Fig. 6.12.
On treatment of DNA fragment with liposomes, the DNA pieces get
encapsulated inside liposomes. These liposomes can adher to cell
membranes and fuse with them to transfer DNA fragments. Thus, the
DNA enters the cell and then to the nucleus. The positively charged
liposomes very efficiently complex with DNA, bind to cells and
transfer DNA rapidly.

Lipofection is a very efficient technique and is used for the transfer of

genes to bacterial, animal and plant cells.

Method # 5. Transduction:
Sometimes, the foreign DNA can be packed inside animal viruses.
These viruses can naturally infect the cells and introduce the DNA into
host cells. The transfer of DNA by this approach is referred to as
transduction.

Method # 6. Direct Transfer of DNA:

It is possible to directly transfer the DNA into the cell nucleus.
Microinjection and particle bombardment are the two techniques
commonly used for this purpose.

Microinjection:
DNA transfer by microinjection is generally used for the cultured cells.
This technique is also useful to introduce DNA into large cells such as
oocytes, eggs and the cells of early embryos. The term transfection is
used for the transfer DNA into eukaryotic cells.

Transgenic Organisms
Transgenic organisms contain foreign DNA that has been introduced using
biotechnology. Foreign DNA (the transgene) is defined here as DNA from another
species, or else recombinant DNA from the same species that has been manipulated in
the laboratory then reintroduced. The terms transgenic organism and genetically
modified organism (GMO) are generally synonymous. The process of creating
transgenic organisms or cells to be come whole organisms with a permanent change to
their germline has been called either transformation or transfection. (Unfortunately,
both words have alternate meanings. Transformation also refers to the process of
mammalian cell becoming cancerous, while transfection also refers to the process of
introducing DNA into cells in culture, either bacterial or eukaryote, for a temporary use,
not germ line changes.) Transgenic organisms are important research tools, and are
often used when exploring a gene’s function. Transgenesis is also related to the medical
practice of gene therapy, in which DNA is transferred into a patient’s cells to treat
disease. Transgenic organisms are widespread in agriculture. Approximately 90% of
canola, cotton, corn, soybean, and sugar beets grown in North America are transgenic.
No other transgenic livestock or crops (except some squash, papaya, and alfalfa) are
currently produced in North America.

To make a transgenic cell, DNA must first be transferred across the cell membrane,
(and, if present, across the cell wall), without destroying the cell. In some
cases, naked DNA (meaning plasmid or linear DNA that is not bound to any type
of carrier) may be transferred into the cell by adding DNA to the medium and
temporarily increasing the porosity of the membrane, for example by electroporation.
When working with larger cells, naked DNA can also be microinjected into a cell using
a specialized needle. Other methods use vectors to transport DNA across the
membrane. Vectors for transformation/transfection include vesicles made of lipids or
other polymers that surround DNA; various types of particles that carry DNA on their
surface; and infectious viruses and bacteria that naturally transfer their own DNA into a
host cell, but which have been engineered to transfer any DNA molecule of interest.
Usually the foreign DNA is a complete expression unit that includes its own cis-
regulators (e.g. promoter) as well as the gene that is to be transcribed.

When the objective of an experiment is to produce a stable (i.e. heritable) transgenic

eukaryote, the foreign DNA must be incorporated into the host’s chromosomes. For this
to occur, the foreign DNA must enter the host’s nucleus, and recombine with one of the
host’s chromatids. In some species, the foreign DNA is inserted at a random location in
a chromatid, probably wherever strand breakage and non-homologous end joining
happen to occur. In other species, the foreign DNA can be targeted to a particular
locus, by flanking the foreign DNA with DNA that is homologous to the host’s DNA at
that locus. The foreign DNA is then incorporated into the host’s chromosomes through
homologous recombination.

Furthermore, to produce multicellular organisms in which all cells are transgenic and
the transgene is stably inherited, the cell that was originally transformed must be either
a gamete or must develop into tissues that produce gametes. Transgenic gametes can
eventually be mated to produce homozygous, transgenic offspring. The presence of the
transgene in the offspring is typically confirmed using PCR or Southern blotting, and the
expression of the transgene can be measured using reverse-transcription PCR (RT-
PCR), RNA blotting, and Western (protein blotting).

The rate of transcription of a transgene is highly dependent on the state of the

chromatin into which it is inserted (i.e. position effects), as well as other factors.
Therefore, researchers often generate several independently transformed/transfected
lines with the same transgene, and then screen for the lines with the highest
expression. It is also good practice to clone and sequence the transgenic locus from a
newly generated transgenic organism, since errors (truncations, rearrangements, and
other mutations) can be introduced during transformation/transfection.

Biotechnology In Plant and Agriculture

 Agricultural biotechnology is the term used in crop and livestock improvement
through biotechnology tools. Biotechnology encompasses a number of tools and
elements of conventional breeding techniques, bioinformatics, microbiology,
molecular genetics, biochemistry, plant physiology, and molecular biology. The
biotechnological tools that are important for agricultural biotechnology include
conventional plant breeding, tissue culture and micropropagation, molecular
breeding or marker-assisted selection, and genetic engineering and GM crops.
In this chapter, readers would learn about the role of biotechnology in crop
improvement and the major applications of the field.

Biotechnology has given a new dimension to scientific innovations, offering

efficient and cost-effective means to produce a diverse array of novel, value-
added products and tools. The present and future focus is on continuing
improvement of agronomic traits such as yield and abiotic stress resistance in
addition to the biotic stress tolerance of the present generation, crop plants as
biomass feedstocks for biofuels and “biosynthetics,” value-added output traits
such as improved nutrition and food functionality, and plants as production
factories for therapeutics and industrial products. From a consumer
perspective, the focus on value-added traits, especially improved nutrition, is of
greatest interest. Both traditional plant breeding and biotechnology-based
techniques are needed to produce plants with the desired quality traits.
Continuing improvements in molecular and genomic technologies are
contributing to the acceleration of product development.

With almost 870 million people estimated to suffer from chronic hunger
worldwide, undernourishment represents a major problem that severely affects
people in developing countries. In addition to undernourishment, micronutrient
deficiency alone can be a cause of serious illness and death. Large portions of
the world population rely on a single, starch-rich crop as their primary energy
source, and these staple crops are generally not rich sources of micronutrients.
As a result, physical and mental health problems related to micronutrient
deficiencies are estimated to affect around two billion people worldwide. The
situation is expected to get worse in parallel with the expanding world
population. Improving the nutritional quality of staple crops seems to be an
effective and straightforward solution to the problem.

Biotechnological Methods Used In Crop Production

In the 1970s, a series of complementary advances in the field of molecular biology
provided scientists with the ability to readily move DNA between more distantly related
organisms. Today, this recombinant DNA technology has reached a stage where
scientists can take a piece of DNA containing one or more specific genes from nearly
any organism, including plants, animals, bacteria, or viruses, and introduce it into a
specific crop species. The application of recombinant DNA technology frequently has
been referred to as genetic engineering. An organism that has been modified,
or transformed, using modern techniques of genetic exchange is commonly referred to
as a genetically-modified organism (GMO). However, the offspring of any traditional
cross between two organisms also are "genetically modified" relative to the genotype of
either of the contributing parents. Plants that have been genetically modified using
recombinant DNA technology to introduce a gene from either the same or a different
species also are known as transgenic plants and the specific gene transferred is
known as a transgene. Not all GMOs involve the use of cross-species genetic
exchange; recombinant DNA technology also can be used to transfer a gene between
different varieties of the same species or to modify the expression of one or more of a
given plant's own genes, e.g., to amplify the expression of a gene for disease
resistance.

The application of recombinant DNA technology to facilitate genetic exchange in crops

has several advantages over traditional breeding methods. The exchange is far more
precise because only a single (or at most, a few), specific gene that has been identified
as providing a useful trait is being transferred to the recipient plant. As a result, there is
no inclusion of ancillary, unwanted traits that need to be eliminated in subsequent
generations, as often happens with traditional plant breeding. Application of
recombinant DNA technology to plant breeding also allows more rapid development of
varieties containing new and desirable traits. Further, the specific gene being
transferred is known so the genetic change taking place to bring about the desired trait
also is known, which often is not the case with traditional breeding methods where the
fundamental basis of the trait being introduced may not be known at all. Finally, the
ability to transfer genes from any other plant or other organism into a chosen recipient
means that the entire span of genetic capabilities available among all biological
organisms has the potential to be genetically transferred or used in any other organism.
This markedly expands the range of useful traits that ultimately can be applied to the
development of new crop varieties. As a hypothetical example, if the genes that allow
certain bacteria to tolerate high external levels of salt can serve the same purpose when
transferred into crops such as potatoes, wheat, or rice, then the production of such
improved food crops on marginally saline lands may be possible. Given that the
acreage of such saline lands is estimated to be equal to 20 to 25% of the land currently
under cultivation world wide, this would be a significant contribution toward global food
security.

Two primary methods currently exist for introducing transgenic DNA into plant genomes
in a functional manner. For plants known as dicots (broad-leaved plants such as
soybean, tomato, and cotton), transformation is usually brought about by use of a
bacterium, Agro-bacterium tumefaciens. Agrobacterium naturally infects a wide range of
plants and it does so by inserting some of its own DNA directly into the DNA of the
plant. By taking out the undesired traits associated with Agrobacterium infection and
inserting a gene(s) of interest into the Agrobacterium DNA that will ultimately be
incorporated into the plant's DNA, any desired gene can be transferred into a dicot's
DNA following bacterial infection. The cells containing the new gene subsequently can
be identified and grown using plant cell culture technology into a whole plant that now
contains the new transgene incorporated into its DNA. Plants known as monocots
(grass species such as maize, wheat, and rice) are not readily infected
by Agrobacterium so the external DNA that is to be transferred into the plant's genome
is coated on the surface of small tungsten balls and the balls are physically shot into
plant cells. Some of the DNA comes off of the balls and is incorporated into the DNA of
the recipient plant. Those cells can also be identified and grown into a whole plant that
contains the foreign DNA.

The ability to easily incorporate genetic material from virtually any organism into many
different crop plants has reached the stage of commercial applicability. About 50% of
the maize, soybeans, and cotton grown in the United States in 1999 had been modified
using recombinant technologies. The major technical limitation on the application of
recombinant DNA technology to improving plants is insufficient understanding of exactly
which genes control agriculturally important traits and how they act to do so.

The study of genes involves the rapidly developing field of genomics, which refers to
determining the DNA sequence and identifying the location and function of all the genes
in an organism. It appears that many traits are conserved between species, i.e., the
same gene confers the same trait in different species. Thus, a gene for salt tolerance in
bacteria may confer salt tolerance if it is transferred and expressed in rice or wheat. The
advent of large scale sequencing of entire genomes of organisms as diverse as
bacteria, fungi, plants, and animals, is leading to the identification of the complete
complement of genes found in many different organisms. This is dramatically enhancing
the rate at which an understanding of the function of different genes is being achieved.
From the standpoint of agricultural biotechnology, advances in gen-omics will lead to a
rapid increase in the number of useful traits that will be available to enhance crop plants
in the future.

Biofertilizer
A biofertilizer is a substance which contains living micro-organisms which, when applied to seeds,
plant surfaces, or soil, colonize the rhizosphere or the interior of the plant and promotes growth by
increasing the supply or availability of primary nutrients to the host plant.

Biopesticide
Biopesticides, a contraction of 'biological pesticides', include several types of pest management
intervention: through predatory, parasitic, or chemical relationships. The term has been associated
historically with [biological control] – and by implication – the manipulation of living organisms.

 Biotechnology In Animal Production

Animal production biotechnology can be defined as the application of scientific and engineering
principles to the processing or production of materials by animals or aquatic species to provide goods
and services for the well being of human population. Examples of animal biotechnology include
generation of transgenic animals or transgenic fish (animals or fish with one or more genes introduced
by human intervention), using gene knockout technology to generate animals in which a specific gene
has been inactivated, production of nearly identical animals by somatic cell nuclear transfer (also
referred to as clones), or production of infertile aquatic species. However, the only alternative way to
improvement or increase the animal production performance is through application of assisted
reproductive techniques (ART) in the farm practices. The techniques are such as semen
cryopreservation, artificial insemination (AI), oestrus synchronisation and superovulation, laparoscopic
ovum pick-up (LOPU), in vitro maturation, fertilisation and culture (IVMFC), intracytoplasmic sperm
injection (ICSI), embryo sexing, embryo/oocyte cryopreservation, cloning, stem cell, embryo transfer,
ultrasonography (pregnancy diagnosis) and radioimmunoassay (RIA).
genetic engineering, scientists can precisely transfer a beneficial gene (for disease resistance,
for example) from one animal species to another. Cloning technology is a type of breeding
technology to produce an exact genetic copy of an animal – usually a high quality animal with
desirable breeding traits.

Human hemoglobin can be produced in microbial and mammalian organisms using

many different expression systems. It is anticipated that recombinant hemoglobins (or
globin genes) will have many applications including as an infectious agent-free,
inexpensive raw material for a red blood cell substitute, as a vehicle for expression or
delivery of other biomolecules, and in gene therapy of inherited hemoglobinopathies.
Recombinant expression, combined with site-directed mutagenesis, is facilitating the
modification of the functional properties of hemoglobin. Although a functional
hemoglobin molecule can be derived using many of the known expression systems, the
choice of the production system for manufacturing depends on the scale, acceptable
cost, and the associated environmental impact of the various processes. While the
efficient bacterial production systems yield a modified, "surrogate" hemoglobin, the
transgenic animal-derived product is virtually identical to the human erythrocyte-
derived hemoglobin. Both the microbial fermentation, and the mammalian transgenic
systems can be geared to produce the enormous quantities of hemoglobin expected to
be required to meet the anticipated demand for a successful blood substitute in the
future.

Cell culture is the process by which human, animal, or insect cells are
grown in a favorable artificial environment. The cells may be derived
from multicellular eukaryotes, already established cell lines, or
established cell strains. In the mid-1900s animal cell culture became a
common laboratory technique, but the concept of maintaining live cell
lines separated from their original tissue source was discovered in the
19th century. Animal cell culture is now one of the major tools used in
the life sciences in areas of research that have a potential for
economic value and commercialization. The development of basic
culture media has enabled scientists to work with a wide variety of
cells under controlled conditions; this has played an important role in
advancing our understanding of cell growth and differentiation,
identification of growth factors, and understanding of mechanisms
underlying the normal functions of various cell types. New
technologies have also been applied to investigate high cell density
bioreactor and culture conditions.
Many products of biotechnology (such as viral vaccines) are
fundamentally dependent on mass culturing of animal cell lines.
Although many simpler proteins are being produced
using rDNA in bacterial cultures, more complex proteins that are
glycosylated (carbohydrate-modified) currently have to be made in
animal cells. At present, cell culture research is aimed at investigating
the influence of culture conditions on viability, productivity, and the
constancy of post-translational modifications such as glycosylation,
which are important for biological activity of recombinant proteins.
Biologicals produced by recombinant DNA (rDNA) technology in
animal cell cultures include anticancer agents, enzymes,
immunobiologicals (interleukins, lymphokines, monoclonal antibodies),
and hormones.