GOVT.

ARTS & SCIENCE COLLEGE

RATLAM

Topic : GENETIC ENGINEERING

Submitted to: Submitted by:

Ms Kanushri Ranawat Shivangi Soni
M.SC FINAL
BIOTECHNOLOGY
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 INDEX

• Introduction
• Steps involved in Genetic Engineering
• Tools used in Genetic Engineering
 Gene of interest (DNA insert)
 Restriction Enzymes
 Vector
 Host cell
• Gene transfer methods
• In vitro production of Insulin
• Transgenic plant
• Transgenic animal
• Application
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 HISTORY

Father of Genetic Engineering is Paul Berg. He was the

first who developed recombinant DNA technology.

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 THE BEGINNING

• The first genetically modified

animal was mouse created in
1973 by Rudolf Jaenisch.
• In 1993, an antibiotic resistant
gene was inserted in tobacco
plant, leading to first genetically
modified plant.
• In 1978, the technology was
commercialized with the
production of insulin.
• In 1994, first genetically modified
food Tomato was made.
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 DEFINATION

The change in genetic make up of living cells by

inserting desired gene through a vector in called
genetic engineering (GE).

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 TERMS

• Gene: The gene is small piece of DNA that

encodes for a specific protein.
• Recombinant DNA: The DNA formed by joining
DNA segment of two different organism.
• Recombinant DNA technology: The technique by
which gene of interest is transferred to the host.
• Genetically modified organism: The organism
whose genetic make up is altered/changed using
rDNA technology.

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 PROCEDURE
• Isolation of desired DNA fragment(gene of interest)
with the help of restriction enzymes.
• Isolation of DNA vector.
• Construction of rDNA. In this gene of interest is
inserted into the vector.
• Introduction of vector containing recombinant into
the host cell.
• Multiplication of Host cells containing recombinant
DNA.
• Expression of cloned gene.
• Selection of Recombinant cells.
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Selection of transformed cell

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 TOOLS USED IN GENETIC ENGINEERING

1. Restriction Endonuclease(RE):
These are the enzyme which cleaves the DNA from
particular sequence.
The sequence from where it cleaves the DNA is called
as recognition sequence. Recognition site can be 4 to
8 bp long.
It breaks the nucleotide bond of base pair.

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 TYPES OF RESTRICTION ENDONUCLEASE

• Type I : Made up of three non-identical subunit.

Require ATP, mg2+ for activation. They cleave the
DNA 1000 bp away from the recognition.
• Type II : Require only mg2+. Made of two identical
subunit. Cleaves DNA from recognition site. These
are widely used enzyme. More than 300 enzyme are
discovered.
• Type III : Cleave 26 bp away from recognition site.

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 CLEAVAGE TYPE

Blunt end ->

Sticky end ->

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 EXAMPLE OF RESTRICTION ENDONUCLEASE

Restriction Source(organism) Recognition

endonuclease sequence
AluI Arthrobacter luteus AG/CT
TC/GA
BamHI Bacillus G/GATCC
amyloliquefaciens CCTAG/G
EcoRI Escherichia coli G/AATTC
CTTAA/G
HindII H. influenza A/AGCTT
TTCGA/A
TaqI Thermus aquaticus T/CGA
AGC/T
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 2) Gene library

The gene of interest (DNA fragment) is stored

in gene library. There are two gene library
available.
Genomic library : A collection clones contain
all DNA segments of the genome of an
organism is called Genomic library.
cDNA library : A collection of clones each of
which carries a cDNA of an organism is called
cDNA.
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 3) VECTOR
A vector is a DNA molecule that
has the ability to replicate
autonomously in an host cell and
into which the DNA fragment to
be cloned.
Any extra chromosomal small
genome/DNA, self replicating
e.G :- Plasmid(pBR322,
pUC18/19), Phage(λ phage,
phage M13), Cosmid, Phasmid,
BAC, YAC.
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 PROPERTIES OF GOOD VECTOR

• Should have origin of

replication.
• Should be less than
10kb in size.
• Easy to isolate and
purify.
• Easily introduced to
host cell.
• Should contain unique
target site for many RE.

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 4) HOST CELL

Host cell are the organism in which rDNA are to be

transformed. E.g:- The best example for host cell is
E. coli.
Properties of good vector:
1. Easy to transform.
2. Support the replication of rDNA
3. Lack active restriction enzyme

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 GENE TRANSFER METHOD

The transfer of rDNA into a bacterial cell or plant cell

or animal cell is called gene transfer.
The host cell contain an rDNA is known as transformed
cell or recombinant.
The rDNA in cell replicates independently of the
chromosomal DNA of cell.
The desired foreign gene present in the rDNA express
its characters in host cell.

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 METHODS

In prokaryotes
• Transformation
• Transduction
• Electroporation
In eukaryotes
• Tranfection
• Electroporation
• Ultrasonication
• Bombardment
• Microinjection
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 In vitro PRODUCTION OF INSULIN

• Isolation of gene
responsible for insulin
production
• Isolation of plasmid
• Gene insertion in
plasmid
• Introduction of rDNA in
host
• Multiplication of
recombinant host cell.

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 TRANSGENIC PLANT
The plant that have been
genetically engineered.
They are :-
• Herbicide resistance
• Insect resistance
• Virus resistance
• Improved storage
• Altered flower color
• Environmental stress
resistance
• Improved nutritional
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quality
 E.g. BT COTTON

• Bt cotton is genetically
modified organism
(GMO)cotton variety, which
produce insecticide to
bollworm.

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 TRANSGENIC ANIMAL

Genetically manipulated
animal having an
introduced gene are
called transgenic
animal.
Transgenic mice, sheep,
cattle, goats, pigs,
poultry and fishes have
been developed by
using GE.
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 E.g. GLOFISH

The glow fish is a

genetically modified
fluorescent fish.
Done by
bioluminescence, which
are found in fireflies or
lightning bugs.
A enzyme called as
luciferase is taken from
firefly and is inserted in
fish which make them
to glow.

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 APPLICATION
• Agriculture
Improved crops
High yield
Resistant
High nutritional value
Long storage
• Medicine
Production of insulin and human growth hormone
• Animal husbandry
High milk production
High yield of wool

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Thank you…

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