CERTIFICATE

This is to certify that the seminar report on “Genetic Engineering” has

been submitted by Dinesh Kumar Meher bearing Roll number ****** in
partial fulfillment of the requirements for the award of the Degree of
Master of Computer application.

Faculty in-charge,PG programme Associate Professor & HOD

Dept. of CSE &A Dept. of CSE &A
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ACKNOWLEDGEMENTS

I deemed it a great pleasure and opportunity to dedicate the following

few lines to some persons who enabled me to complete the Seminar.

I express my obligations to ******* HOD, department of Computer

Science Engineering & Applications for making arrangement for the Seminar.

I really indebted to the faculty members of the department for their

suggestions, motivations & invaluable guidance right from scarp up to the
completion of the Seminar. Their contributions show in many subtle ways and
were indeed instrumental in achieving the final result.
.

Dinesh Kumar Meher

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ABSTRACT

Genetic engineering, also called recombinant DNA technology,

involves the group of techniques used to cut up and join together genetic
material, especially DNA from different biological species, and to introduce the
resulting hybrid DNA into an organism in order to form new combinations of
heritable genetic material. These achievements led to concerns in the scientific
community about potential risks from genetic engineering. To address these
concerns, a meeting was held in 1974 at the Asilomar Conference Center in
California. The conference was a milestone in the development of social
awareness and of public responsibility among scientists. A few of the
innovators of the new technology realized its commercial potential and
established private biotechnology companies. One of the first to do this was
Boyer who founded Genentech Inc. The company developed the production of
human insulin in bacteria. Genetically engineered human insulin has provided a
reliable, expandable, and constant supply for diabetics around the world. The
methods for genetically engineering bacteria, plants, and animals are discussed,
as well as the arguments, pro and con, for GM plant and animal products.
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LIST OF FIGURES

Figure 5.1 Making Transgenic Mice 8

Figure 5.2.1 Restriction Endonucleases 11

Figure 5.2.1 Recombinant DNA 12

Figure 6.1 DOLLY - 1960 14

Figure 6.2 Cloning of a EWE 15

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CONTENTS

CERTIFICATE I

ACKNOWLEDGEMENTS II

ABSTRACT III

LIST OF FIGURES IV

CONTENTS V

1 Introduction 1

2 What is Genetic Engineering 2

3 History 3-4

4 Techniques of Genetic Engineering 5

5 Prospects for Genetic Engineering 6-13

5.1 Transgenic Engineering 6-8

5.2 Cloning 9-13

6 DOLLY - 1960 14-15

7 Concerns in Cloning 16

8 Dangers of Genetic Engineering 17-18

9 Advantages 19

10 Conclusion 20

11 References 21
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Introduction

What is genetics?

Genetics is the scientific study of genes, i.e. variations in the characteristics

-- resemblances and differences -- of organisms and how these
characteristics are inherited from generation to generation. Modern genetics
is as much concerned with the organism level of this process as it is with the
cellular and molecular level.

What are genes?

A gene is a fuzzy concept which depends upon who is using the term and in
what context. For the earliest geneticists, genes were fairly distinct traits or
characteristics which could be observed in the whole organism. For the
modern molecular biologist or molecular geneticist a more chemical
definition of a gene is used which brings in a number of additional concepts.
The molecular gene is a definite sequence of bases in the DNA chain which
together code for the production of a particular protein.
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What is genetic engineering?

Engineering is the technological manipulation of the objects of the natural
world in a way that is perceived to be beneficial to people. Traditionally we
used the word in the context of inanimate nature: bridges, railways and
machines etc. But the term can be used and is used in the context of biology,
namely for bioengineering, i.e. modifying or manipulating living organisms.
Another term used in place of the term 'genetic engineering' is
'biotechnology'.

Definition of Genetic Engineering

“Genetic engineering is the technology for modifying the genetic information
in a plant, animal or human in order to produce some desired trait or
characteristic”
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History of Genetic Engineering

Modern genetic engineering began in 1973 when Herbert Boyer and Stanley
Cohen used enzymes to cut a bacteria plasmid and insert another strand of
DNA in the gap. Both bits of DNA were from the same type of bacteria, but
this milestone, the invention of recombinant DNA technology, offered a
window into the previously impossible -- the mixing of traits between totally
dissimilar organisms. To prove that this was possible, Cohen and Boyer used
the same process to put a bit of frog DNA into bacteria.

Since 1973, this technology has been made more controllable by the
discovery of new enzymes to cut the DNA differently and by mapping the
genetic code of different organisms. Now that we have a better idea of what
part of the genetic code does what, we have been able to make bacteria that
produce human insulin for diabetics (previously came from livestock), as
well as EPO for people on kidney dialysis (previously came from urine of
people in third world countries with ringworm).

In 1990, a young child with an extremely poor immune system

received genetic therapy. Some of her white blood cells were genetically
manipulated and re-introduced into her bloodstream while she watched
Sesame Street. These new cells have taken over for the original, weak white
cells, and her immune system now works properly. Although relatively few
people have had their cells genetically altered, these advances have made the
prospect of mainstream genetic medicine seem more likely.
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As of late summer of 1998, scientists are able to add simple traits to

organisms. They cannot create custom-made animals. They cannot always
predict how traits will interact. Before phenomenally new advances can be
made, scientists have to learn how to affect cells' DNA with pin-point
accuracy, without affecting other traits. Advances like genetic correction for
nearsightedness are a long way off. The power of science is limited to
knowledge about genetics, gene locations, and trait interactions, but as
knowledge grows, so will scientists' abilities to manipulate life.
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Techniques of Genetic Engineering

Recombinant DNA techniques use biological vectors like plasmids
and viruses to carry foreign genes into cells. Plasmids are small circular
pieces of genetic material found in bacteria that have the ability to cross
species boundaries. The circles can be broken and new genetic material
added to them. Plasmids augmented with new genetic material can move
across microbial cell boundaries and place the new genetic material next to
the bacterium's own genes. Often the bacteria will take up the gene and
begin to produce the protein for which the gene codes. Where the new gene
codes for insulin, for example, the bacterium will begin to produce insulin
along with its other gene products. A large vat of bacteria engineered to
produce insulin can then become a sort of pharmaceutical factory.
Viruses can also act as vectors in genetic engineering. Viruses are
infectious particles that contain genetic material to which a new gene can be
added. The virus can carry the new gene into a recipient cell in the process of
infecting that cell. The virus can also be disabled so that while it can carry a
new gene into a cell, it cannot redirect the cell's genetic machines to make
thousands of copies of itself.

Transformation: - Transformation is a process by which a cell takes up

naked DNA fragment from the environment, incorporates it into its own
chromosomal DNA.
Transduction: - Transfer of DNA from one organism to another through a
bacteriophage is called transduction.
Examples:-
1. Transduction from bacterium to bacterium.

2. Transduction from bacteria to Human cells.

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Prospects for Genetic Engineering

Transgenic Engineering:-
Putting genetic information from one type of plant or animal into another
Plants:-

Transgenic plants possess a gene or genes that have been transferred

from a different species. Although DNA of another species can be integrated in
a plant genome by natural processes, the term "transgenic plants" refers to
plants created in a laboratory using recombinant DNA technology. The aim is
to design plants with specific characteristics by artificial insertion of genes
from other species or sometimes entirely different kingdoms.

The intentional creation of transgenic plants by laboratory based

recombinant DNA methods is more recent (from the mid-70s on) and has
been a controversial development in the field of biotechnology opposed
vigorously by many NGOs, and several governments, particularly within the
European Community. These transgenic recombinant plants ( biotech crops,
modern transgenics) are transforming agriculture in those regions that have
allowed farmers to adopt them, and the area sown to these crops has
continued to grow globally in every years since their first introduction in
1996.

Transgenic recombinant plants are generated in a laboratory by

adding one or more genes to a plant's genome,and the techniques frequently
called transformation.
Transformation is usually achieved using gold particle bombardment or
through the process of Horizontal gene transfer using a soil bacterium,
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Agrobacterium tumefaciens, carrying an engineered plasmid vector, or

carrier of selected extra genes.
Transgenic recombinant plants are identified as a class of genetically
modified organism(GMO); usually only transgenic plants created by direct
DNA manipulation are given much attention in public discussions.

Transgenic Animal:-

Transgenic mice contain additional foreign DNA in every cell allowing

them to be used to study gene function or regulation and to model human
diseases. Transgenic mouse contains additional, artificially-introduced
genetic material in every cell. This often confers a gain of function, for
example the mouse may produce a new protein, but a loss of function may
occur if the integrated DNA interrupts another gene. A transgenic mouse a
very useful system for studying mammalian gene function and regulation
because analysis is carried out on the whole organism.

Transgenic mice are also used to model human diseases that involve
the over expression or misexpression of a particular protein.
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Making Transgenic Mice:-

Figure 5.1
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Cloning:-
Making genetic copies of an existing human plant or
animal. Asexual breeding in plants & lower animals.

Human:-

Human cloning is the creation of a genetically identical copy of a

human being, human cell, or human tissue. The term is generally used to
refer to artificial human cloning; human clones in the form of identical twins
are commonplace, with their cloning part of the natural process of
reproduction.

Although genes influence behavior and cognition, "genetically

identical" does not mean altogether identical; identical twins, despite being
natural human clones with nearly identical DNA, are separate people, with
separate experiences and personalities. The relationship between an
"original" and a clone is rather like that between identical triplets raised
apart; they share nearly all of the same DNA, but little of the same
environment. A lively scientific debate on this topic occurred in the journal
Nature in 1997. Ultimately, the question of how similar an original and a
clone would be boils down to how much of personality is determined by
genetics, an area still under active scientific investigation.
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Steps in Cloning a Gene :-

Let us walk through the basic steps for cloning a gene, a process by
which a gene of interest can be replicated many times over. Let us pretend that
we are going to genetically engineer E. coli cells to glow in the dark, a
characteristic that they do not naturally possess.

1. Isolate DNA of interest – first we need to identify the genes or genes

that we are interested in, the target DNA. If we want our E. coli cells to
glow in the dark, we need to find an organism that possesses this trait
and identify the gene or genes responsible for the trait. The green
fluorescent protein (GFP) commonly used as an expression marker in
molecular techniques was originally isolated from jellyfish.In cloning a
gene it is helpful to use a cloning vector, typically a plasmid or virus,
capable of independent replication that will stably carry the target DNA
from one location to another. Plasmid vectors are available from both
bacteria and yeast.
2. Cut DNA with restriction endonucleases – once the target and vector

DNA have been identified, both types of DNA are cut using restriction
endonucleases. These enzymes recognize short sequences of DNA that
are 4-8 bp long. The enzymes are widespread in both bacteria and
archaea, with each enzyme recognizing a specific inverted repeat
sequence that is palindromic (reads the same on each DNA strand, in the
5’ to 3’ direction).
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Restriction Endonucleases.
Figure 5.2.1
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While some restriction endonucleases cut straight across the DNA (i.e.
blunt cut), many make staggered cuts, producing a very short region of
single-stranded DNA on each strand. These single-stranded regions are
referred to as “sticky ends,” and are invaluable in molecular cloning
since the unpaired bases will recombine with any DNA having the
complementary base sequence.

3. Combine target and vector DNA – after both types of DNA have been

cleaved by the same restriction endonuclease, the two types of DNA are
combined together with the addition of DNA ligase, an enzyme that
repairs the covalent bonds on the sugar-phosphate backbone of the DNA.
This results in the creation of recombinant DNA, DNA molecules that
contain the DNA from two or more sources, also known as chimeras.

Figure 5.2.2
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4. Introduce recombined molecule into host cell – once the target DNA has

been stably combined with vector DNA, the recombinant DNA must be
introduced into a host cell, in order for the genes to be replicated or
expressed. There are different methods for introducing the recombinant
DNA, largely depending upon the complexity of the host organism. In
the case of bacteria, transformation is often the easiest method, using
competent cells to pick up the recombinant DNA molecules.
Alternatively, electroporation can be used, where the cells are exposed
to a brief pulse of high –voltage electricity causing the plasma membrane
to become temporarily permeable to DNA passage.While some cells will
acquire recombinant DNA with the appropriate configuration (i.e. target
DNA combined with vector DNA), the method also will yield cells
carrying recombinant DNA with alternate DNA combinations (i.e.
plasmid DNA combining with another plasmid DNA molecule or target
DNA attached to more target DNA). The mixture is referred to as
a genomic library and must be screened to select the appropriate clone.
If random fragments of DNA were originally used (instead of isolation of
the appropriate target DNA genes), the process is referred to as shotgun
cloning and can yield thousands or tens of thousands of clones to be
screened.
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DOLLY-1960
Dolly (July 5, 1996 – February 14, 2003), a female sheep or ewe, was
the first animal to be cloned from an adult somatic cell, using the process
of nuclear transfer. She was cloned by Ian Wilmut, Keith Campbell and
colleagues at the Roslin Institute in Edinburgh, Scotland. Her birth was
announced on February 22, 1997 and she lived until the age of six.

The cell used as the donor for the cloning of Dolly was taken from a
mammary gland, and the production of a healthy clone therefore proved that
a cell taken from a specific body part could recreate a whole individual.
More specifically, the production of Dolly showed that mature
differentiated somatic cells in an adult animal's body could under some
circumstances revert back to an undifferentiated pluripotent form and then
develop into any part of an animal. As Dolly was cloned from part of a
mammary gland, she was named after the famously curvaceous country
western singer Dolly Parton.

Figure 6.1
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Figure 6.2

Cloning since Dolly:-

Cloning of this sort has now been done on cattle, pigs and mice
also. The success rate has improved considerably.Cloning humans
begins to show up in science fiction in 1970s.This is now a realistic
possibility.
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Concerns in Cloning
 The success rate from adult animal cells is still rather low.
 This would be unacceptable for cloning humans in most societies.
 The evidence suggests that the clones which survive are still not
right.
 The genetic program has probably not been completely reset.
 We still don’t understand what we are doing in cloning from adult
cells.

Advantages of Cloning:-

With an adult plant or animal, the breeder knows what its traits are; this
is not the case with fetal cell cloning.
Cloning allows making a genetically identical copy of the desired plant
or animal.
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Dangers related to Genetic

Engineering

 Imprecise Technology—a genetic engineer moves genes from

one organism to another. A gene can be cut precisely from the DNA
of an organism, but the insertion into the DNA of the target
organism is basically random. As a consequence, there is a risk that
it may disrupt the functioning of other genes essential to the life of
that organism.
 Side Effects—Genetic engineering is like performing heart surgery
with a shovel. Scientists do not yet understand living systems
completely enough to perform DNA surgery without creating
mutations which could be harmful to the environment and our health.
They are experimenting with very delicate, yet powerful forces of
nature, without full knowledge of the repercussions.
 Widespread Crop Failure - Genetic engineers intend to profit

by patenting genetically engineered seeds. This means that, when a

farmer plants genetically engineered seeds, all the seeds have
identical genetic structure. As a result, if a fungus, a virus, or a pest
develops which can attack this particular crop, there could be
widespread crop failure.
 Threatens Our Entire Food Supply—Insects, birds, and wind
can carry genetically altered seeds into neighboring fields and beyond.
Pollen from transgenic plants can cross-pollinate with genetically
natural crops and wild relatives. All crops, organic and non-organic, are
vulnerable to contamination from cross-pollinatation.
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 No Long-Term Safety Testing—Genetic engineering uses

material from organisms that have never been part of the human food
supply to change the fundamental nature of the food we eat. Without
long-term testing no one knows if these foods are safe

 Allergic Reactions—Genetic engineering can also produce

unforeseen and unknown allergens in foods.)

 Decreased Nutritional Value—transgenic foods may mislead

consumers with counterfeit freshness. A luscious-looking, bright red
genetically engineered.
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Advantages

1. Creating new types of human beings with advantageous traits.

2. GM animals can generate pharmaceutical proteins.

3. Quicker, more predictable way to generate new cultivars.

4. Sustainable agriculture
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Conclusion
Engineering is the technological manipulation of the objects of the
natural world in a way that is perceived to be beneficial to people. With
this technology we modify the genetic information in a plant, animal or
human in order to produce some desired trait or characteristic which are
very beneficial for us.
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References

 www.ieeeexplore.ieee.org
 www.geneticengineering.org
 www.wikipedia.com/geneticengineering