This document discusses recombinant DNA technology, which involves introducing foreign DNA into a host organism. It describes the basic steps of isolating DNA, cutting it with restriction enzymes, amplifying the gene with PCR, and ligating it into a vector for insertion into the host. The key tools used are restriction enzymes, DNA polymerase for PCR, ligase, and vectors to introduce the recombinant DNA into hosts such as bacteria. The goal is to generate organisms that produce desired proteins or have new traits.

1. Recombinant DNA
Technology

2. Content:-
 Introduction
 Purpose of Recombinant DNA Technology
 Enzymes of Recombinant DNA Technology
 Tools of Recombinant DNA Technology
 Process of recombinant DNA Technology

3. The basic steps of recombinant DNA
technology
Using the bacterial plasmid as cloning
vectors

4. INTRODUCTION TO RECOMBINANT DNA
TECHNOLOGY
A technique mainly used to change the
phenotype of an organism (host) when a
genetically altered vector is introduced
and integrated into the genome of the
organism. So, basically, this process
involves the introduction of a foreign
piece of DNA structure into the genome
which contains our gene of interest. This
gene which is introduced is the
recombinant gene and the technique is
called the recombinant DNA technology.
There are multiple steps, tools and other
specific procedure followed in the
recombinant DNA technology, which is
used for producing artificial DNA to
generate the desired product.

5. Purpose Of Recombinant DNA Technology
mRNA/DNA
CDNA/DNA
fragments
DNA Probe
CDNA bank
DNA BAnk
Cloning vector
(plasmid,
Casmides Virus)
Ligation

6. Plant
cell/protoplast
Culture
Recombinant
DNA
Transformed
cell
Resired product
nif genes, stress and
disease resistant
genes, other genes
Vaccines,
interferon,
Antibiotic,
chemicals,
monoclinal, etc.
Application Of Genetic Engine – RING TECHNIQUES

8. Enzymes Involved In Recombinant DNA Technology

10. 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
Restriction Enzymes
polymerase
Ligase
Vectors
Host Organism

11. Process of Recombinant DNA Technology
The complete process of recombinant DNA
technology includes multiple steps, maintained in a specific
sequence to generate the desired product.
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.

14. AMPLIFICATION OF THE
DNA FRAGMENTS
The polymerase chain reaction (PCR) is a biochemical
technology in molecular biology used to amplify a
single, or a few copies, of a piece of DNA across
several orders of magnitude, generating thousands to
millions of copies of a particular DNA sequence.
LIGATION OF DNA
FRAGMENTS INTOVECTOR

15. Insertion of recombinant DNA
into the host
The host is the final tool of rDNA technology, which consumes
the vector engineered with the desired DNA with the aid of the
enzymes. Insertion of the desired recombinant DNA into the
host organism can be achieved in various ways. This includes–
biolistics or the gene gun, microinjection, alternate heating and
cooling, usage of calcium ions, etc. The successfully transformed
cells or the entities pass the recombinant gene to the offspring.
Downstream Processing

16. Bibliography
 NCERT textbook { class XII}
 WWW.GOOGLE.CO.IN
 WIKIPEDIA
 BYJUS
 VEDANTU
 LIBRARY
 REFFERENCE