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amrit_rj

Reach out to me @
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 PRINCIPLES OF
BIOTECHNOLOGY

GENETIC ENGINEERING

MAINTENANCE OF
STERILE AMBIENCE

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 BIOTECHNOLOGY

Why we prefer Because biotechnology

biotechnology allows us to deliver only
desired set of gene into
over traditional
target organisms not the
breeding undesirable one
methods???

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Origin of replication

● Specific DNA sequence

● Responsible for initiating

replication (making numerous
copies of DNA / RNA)

● When an alien DNA enters, it will

binds to at the “ORI” site, and
initiate multiplying itself inside
host organisms.

● This process can also be called as

cloning.

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 PLASMID

“ Autonomously replicating circular

extra-chromosomal DNA”

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 INTRODUCTION OF
RECOMBINANT DNA
● Stanley Cohen and Herbert Boyer in 1972
isolated the antibiotic resistance gene.

● By cutting a piece of DNA from a plasmid

● The cutting of DNA at specific locations

become possible with the discovery of the
so- called

‘Molecular scissors’ - restriction enzymes’

● The cut piece of DNA then linked with the

plasmid DNA
●
These plasmid DNA acts as vectors to
transfer the piece of DNA attached to it

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 Basic steps of genetic
engineering

Identification of DNA with desirable genes

Introduction of the identified DNA into the host.

Maintenance of introduced DNA in the host and

transfer of the DNA to its progeny.

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 TOOLS OF
RECOMBINANT DNA

Restriction Enzymes

Ligases

TOOLS

Vectors

Host Organisms

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RESTRICTION ENZYMES

“MOLECULAR SCISSORS”

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RESTRICTION ENZYMES

Used to cut DNA with different ends.

There are 2 types of restriction enzymes:

Exonucleases
Cuts the DNA at the ends.

Endonucleases
Cuts the DNA in the middle

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NOMENCLATURE OF
RESTRICTION ENZYMES

EcoRI

E co R I

E - Genus Name - Escherichia

Co - Species - coli
R - Strain- RY13
Order of identification - First endonuclease
isolated

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Endonuclease & Exonuclease

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TYPES OF RESTRICTION ENZYMES

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Endonuclease & Exonuclease

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RESTRICTION SITES

● Restriction enzymes cut at a

specific site on DNA -
RESTRICTION SITE

● Restriction endonuclease
recognizes a specific palindromic
nucleotide sequence in the DNA.

● PALINDROMES - group of letters

that from the same word when
read both forwards and backward.

● Such as “MALAYALAM”

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RESTRICTION ENZYMES

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 DNA LIGASE

The sticky ends produced by the

restriction enzyme are then joined by this
molecular glue - DNA ligase enzyme.

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Representation of Recombinant
DNA Technology

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CLONING VECTORS

● Any DNA molecule that carrier a

gene of interest to be inserted
into the host organisms.

● Plasmids, Bacteriophages,
Anima, Plant, Virus, YACs and
BACs are used as cloning
vectors.

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CLONING VECTORS

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Features required in a
cloning vector

Origin of Replication

Selectable Marker

Restriction Sites

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Features required in a
cloning vector

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Origin of replication

● Specific DNA sequence

● Responsible for initiating

replication (making numerous
copies of DNA / RNA)

● When an alien DNA enters, it will

binds to at the “ORI” site, and
initiate multiplying itself inside
host organisms.

● This process can also be called as

cloning.

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 Selectable markers

● Helps in identifying and

eliminating non- transformants
and selectively permitting the
growth of the transformants.

● Generally, the gene coding

resistance to antibiotics such
as ampicillin, chloramphenicol,
tetracycline or kanamycin etc.,
considered useful selectable
markers for E.coli

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 Cloning Sites

● This is the site where restriction

enzyme cuts the DNA.

● The gene of interest in

incorporated in this region.

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 Vectors for cloning in
plants and Animals

● Agrobacterium tumefaciens , a
pathogen of several dicot plants
is used as vector for plants.

● Its Structure of T- DNA is able to

deliver a piece of DNA to
transform normal plant cells into
tumor

● Direct these tumor cells to

produce the chemical required by
the pathogen.

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 Ti Plasmid

● Tumor inducing (Ti) plasmid of

Agrobacterium tumefaciens has
modified into a cloning vector.

● An extra-chromosomal, double
stranded and self replicating DNA
molecule present in
Agrobacterium tumefaciens.

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 Vectors in Animals

● Retroviruses in animals have the

ability to transform normal cells
into cancerous cells.

● Modified into cloning vector for

animals.

● Bacteriophages can also be used

as cloning vector in animals.

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amrit_rj
Plasmid has been used as vector because:

1 Both its end shows replication

It can move between prokaryotic and

2 eukaryotic cells

It is a extrachromosomal circular DNA

3 having autonomously replicating capability

4 It has antibiotic resistance gene

Plasmid has been used as vector because:

1 Both its end shows replication

It can move between prokaryotic and

2 eukaryotic cells

It is a extrachromosomal circular DNA

3 having autonomously replicating capability

4 It has antibiotic resistance gene

An enzyme catalyzing the removal of nucleotides
from the ends of DNA is:

1 Exonuclease

2 Endonuclease

3 Hind II

4 DNA Ligase
An enzyme catalyzing the removal of nucleotides
from the ends of DNA is:

1 Exonuclease

2 Endonuclease

3 Hind II

4 DNA Ligase
The most extensively used bacteria in genetic
engineering is

1 Bacillus

2 Clostridium

3 Escherichia

4 Salmonella
The most extensively used bacteria in genetic
engineering is

1 Bacillus

2 Clostridium

3 Escherichia

4 Salmonella
Vector for T-DNA is:

1 Agrobacterium tumefaciens

2 Thermus aquaticus

3 Salmonella typhimurium

4 Arabidopsis thaliana
Vector for T-DNA is:

1 Agrobacterium tumefaciens

2 Thermus aquaticus

3 Salmonella typhimurium

4 Arabidopsis thaliana
The source of the restriction enzyme Hind III is:

1 Escherichia coli RY 13

2 Haemophilus influenzae Rd

3 Bacillus subtilis

4 Lactobacillus
The source of the restriction enzyme Hind III is:

1 Escherichia coli RY 13

2 Haemophilus influenzae Rd

3 Bacillus subtilis

4 Lactobacillus
ARAPRO
 amrit_rj

Reach out to me @
amrit_rj amrit.raj@vedantu.com