Recombinant DNA

Technology

A PROJECT REPORT IN BIOLOGY (044)

SUBMITTED IN PARTIAL FULFILLMENT OF
THE REQUIREMENT FOR THE COMPLETION
OF

AISSCE 2022-2023

BY Pulkit Dewangan
AISSCE Roll No: ……………………

Under the supervision of

Ms. Pooja Jha
PGT Biology

The Great India School,

Raipur, Chhattisgarh 1
 CERTIFICATE

This is to certify that Pulkit Dewangan of

class 12 has successfully completed the
project work on Biology for class XII AISSCE
practical examination of the Central Board
of Secondary Education in the year 2022-
2023. It is further certified that this project
is the individual work of the candidate.

Internal Examiner External Examiner

Signature Signature

Rajesh Kumar Agrawal

Principal

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ACKNOWLEDGEMENT

I would like to thank my Biology teacher,

Ms. Pooja Jha for guiding me throughout this
project work. I should also thank our lab
assistant who helped me to line up the
project and helped me with practical work.

A special acknowledgement goes to our

Principal Rajesh Kumar Agrawal who gave
me the golden opportunity of this wonderful
project, which also helped me in doing a lot
of research and I came to know about so
many new things.

I wish to thank my parents as well for their

support and encouragement without which I
could not have completed this project in
the limited time frame.

In the end, I want to thank my friends who

displayed appreciation for my work and
motivated me to continue my work.
 INDEX
Sr No. Heading Page No.

1 Introduction 1

2 DNA Cloning 2

3 Tools Of 3
Recombinant
DNA Technology

4 Process of 4
Recombinant
DNA Technology

5 Application of 5
Recombinant
DNA Technology

6 Applications 6
Of Gene Cloning

7 How does rDNA 7

works

8 Products of rDNA 8

10 Bibliography 10
Introduction

 What is recombinant DNA technology?

 Recombinant DNA technology is the joining together
of DNA molecules from two different species. The
recombined DNA molecule is inserted into a host
organism to produce new genetic combinations that
are of value to science, medicine, agriculture, and
industry.

 When was the recombinant DNA technology was

invented?
 The possibility for recombinant DNA technology
emerged with the discovery of restrictin enzymes in
1968 by Swiss microbiologist Werner Arber.

 How recombinant DNA technology is useful?

 Through recombinant DNA techniques, bacteria have
been created that are capable of synthesizing
human insulin, human growth hormone, alpha
interferon, hepatitis B vaccine, and other medically
useful substances. Recombinant DNA technology also
can be used for gene therapy, in which a normal
gene is introduced into an individual’s genome in
order to repair a mutation that causes a genetic
disease.

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DNA Cloning

 In biology a clone is a group of individual cells or

organisms descended from one progenitor. This
means that the members of a clone are genetically
identical, because cell replication produces
identical daughter cells each time. The use of the
word clone has been extended to recombinant DNA
technology, which has provided scientists with the
ability to produce many copies of a single fragment
of DNA, such as a gene, creating identical copies
that constitute a DNA clone.

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Tools Of Recombinant DNA
Technology

 The enzymes which include 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 remove the nucleotides
from the ends of the strands. The restriction
endonucleases are sequence-specific which are
usually palindrome sequences and cut the DNA at
specific points.

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Process of Recombinant DNA
Technology
 Step-1. Isolation of Genetic Material.
 The first and the initial step in Recombinant DNA
technology is to isolate the desired DNA in its pure
form i.e. free from other macromolecules.
 Step-2.Cutting the gene at the recognition sites.
 The restriction enzymes play a major role in
determining the location at which the desired gene
is inserted into the vector genome. These reactions
are called ‘restriction enzyme digestions’.
 Step-3. Amplifying the gene copies through
Polymerase chain reaction (PCR).
 It is a process to amplify a single copy of DNA into
thousands to millions of copies once the proper gene
of interest has been cut using the restriction
enzymes.
 Step-4. Ligation of DNA Molecules.
 In this step of Ligation, joining of the two pieces – a
cut fragment of DNA and the vector together with
the help of the enzyme DNA ligase.
 Step-5. Insertion of Recombinant DNA Into Host.
 In this step, the recombinant DNA is introduced into
a recipient host cell. This process is termed
as Transformation. Once after the insertion of the
recombinant DNA into the host cell, it
gets multiplied and is expressed in the form of the
manufactured protein under optimal conditions.

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Application of Recombinant DNA
Technology

 DNA technology is also used to detect the presence

of HIV in a person.
 Gene Therapy – It is used as an attempt to correct
the gene defects which give rise to heredity
diseases.
 Clinical diagnosis – ELISA is an example where the
application of recombinant
 Recombinant DNA technology is widely used in
Agriculture to produce genetically-modified
organisms such as Flavr Savr tomatoes, golden rice
rich in proteins, and Bt-cotton to protect the plant
against ball worms and a lot more.
 In the field of medicines, Recombinant DNA
technology is used for the production of Insulin.

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Applications Of Gene Cloning

 Gene Cloning plays an important role in the

medicinal field. It is used in the production of
hormones, vitamins and antibiotics.
 Gene cloning finds its applications in the agricultural
field. Nitrogen fixation is carried out by
cyanobacteria wherein desired genes can be used to
enhance the productivity of crops and improvement
of health. This practice reduces the use of fertilizers
hence chemical-free produce is generated
 It can be applied to the science of identifying and
detecting a clone containing a particular gene which
can be manipulated by growing in a controlled
environment
 It is used in gene therapy where a faulty gene is
replaced by the insertion of a healthy gene. Medical
ailments such as leukaemia and sickle cell anaemia
can be treated with this principle.

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How does rDNA work?

 Recombinant DNA works when the host cell

expresses protein from the recombinant genes.
 A significant amount of recombinant protein will not
be produced by the host unless expression
factors are added. Protein expression depends upon
the gene being surrounded by
a collection of signals which provide instructions for
the transcription and translation
of the gene by the cell. These signals include the
promoter, the ribosome binding
site, and the terminator. Expression vectors, in
which the foreign DNA is inserted,
contain these signals. Signals are species
specific. In the case of E. Coli, these signals must
be E. Coli signals as E. Coli is unlikely to understand
the signals of human promoters and terminators.
 Problems are encountered if the gene contains
introns or contains signals which act
as terminators to a bacterial host. This results in
premature termination, and the recombinant
protein may not be processed correctly, be folded
correctly, or may even be degraded.

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Bibliography

 https://www.britannica.com/science/recombinant-
DNA-technology

 https://byjus.com/biology/recombinant-dna-
technology/

 https://www.rpi.edu/dept/chem-eng/Biotech-
Environ/Projects00/rdna/rdna.html

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