ACKNOWLEDGEMENT

I would like to express my special thanks to my

teacher Mr Manavendra Goswami, who gave
me the opportunity to do this wonderful
project of Biology and also helped me with the
idea.
I came to know so many things I am really
thankful to him.
Secondly,I would also like to thanks my parents
friends who helped me a lot to finalize this
project within the limited time frame.
Arnav Bhatt
XII Biology
Restriction enzyme
A restriction enzyme is a protein that recognizes a
specific, nucleotide sequence and cuts the DNA only
at that specific site, which is known as restriction
site or target sequence.
These enzymes are found in bacteria and provide a
defence mechanism against invading viruses. Inside
a prokaryote, the restriction enzymes selectively cut
up foreign DNA in a process called restriction
digestion. The host DNA is protected by a
modification enzyme that modifies the prokaryotic
DNA and blocks clevage.
Over 3,000 restriction enzymes have been studied in
detail, and more than 600 of these are available
commercially. These enzymes are routinely used for
DNA modification in laboratories, and they are a
vital tool in molecular cloning.

History
The term restriction enzyme originated from the studies
of phage λ, a virus that infects bacteria, and the
phenomenon of host-controlled restriction and
modification of such bacterial phage or bacteriophage.
The phenomenon was first identified in work done in the
laboratories of Salvador Luria, Weigle and Giuseppe
Bertani in the early 1950s.It was found that, for a
bacteriophage λ that can grow well in one strain of
Escherichia coli, for example E. coli C, when grown in
another strain, for example E. coli K, its yields can drop
significantly, by as much as 3-5 orders of magnitude. The
host cell, in this example E. coli K, is known as the
restricting host and appears to have the ability to reduce
the biological activity of the phage λ. If a phage becomes
established in one strain, the ability of that phage to
grow also becomes restricted in other strains hence the
name restriction enzyme.The restriction enzymes
studied by Arber and Meselson were type I restriction
enzymes, which cleave DNA randomly away from the
recognition site.
Recognitation site
Restriction enzymes recognize a specific sequence of
nucleotides and produce a double-stranded cut in the
DNA. This area is know as recognitation site.The
recognition sequences can also be classified by the
number of bases in its recognition site, usually between 4
and 8 bases, and the number of bases in the sequence
will determine how often the site will appear by chance
in any given genome.A recognition sequence is a DNA
sequence to which a structural pattern of a DNA-binding
body exhibits binding specificity.
TYPES
Naturally occurring restriction endonucleases are
categorized into four groups (Types I, II, III and IV )
based on their composition and enzyme cofactor
requirements, the nature of their target sequence,
and the position of their DNA cleavage site relative
to the target sequence.All types of enzymes
recognize specific short DNA sequences and carry
out the endonucleolytic cleavage of DNA to give
specific fragments with terminal 5'-phosphates.
They differ in their recognition sequence, subunit
composition, cleavage position, and cofactor
requirements.

Type
I
enzymes cleave at sites remote from a recognition site;
require both ATP and S-adenosyl-L-methionine to
function; multifunctional protein with both restriction
digestion and methylase activities.

Type II enzymes cleave within or at short specific

distances from a recognition site; most require
magnesium; single function (restriction digestion)
enzymes independent of methylase.

Type III enzymes cleave at sites a short distance from a

recognition site; require ATP ; S-adenosyl-L-methionine
stimulates the reaction but is not required; exist as part
of a complex with a modification methylase.

Type IV enzymes target modified DNA, e.g. methylated,

hydroxymethylated and glucosyl-hydroxymethylated DNA

Type V are artificial restriction enzymes (e.g., the cas9-

gRNA complex from CRISPRs) which utilize guide RNAs to
target specific non-palindromic sequences found on
invading organisms. They can cut DNA of variable length,
provided that a suitable guide RNA is provided. The
flexibility and ease of use of these enzymes make them
promising for future genetic engineering applications.

Nomenclature
Each restriction enzyme is named after the bacteria from
which it is isolated.

Rules
 First capital letter indicates genus of the bacterium
from which enzyme is isolated.
 The second and third small letters indicate species
of the bacterium.
Working
Like all enzymes, a restriction enzyme works by shape-to-
shape matching. When it comes into contact with a DNA
sequence with a shape that matches a part of the
enzyme, called the recognition site, it wraps around the
DNA and causes a break in both strands of the DNA
molecule.
Each restriction enzyme recognises a different and
specific recognition site, or DNA sequence. Recognition
sites are usually only short - 4-8 nucleotides.

Applications
Isolated restriction enzymes are used to manipulate
DNA for different scientific applications.
They are used to assist insertion of genes into
plasmid vectors during gene cloning and protein
production experiments.
Restriction enzymes can be used to distinguish gene
alleles by specifically recognizing single base
changes in DNA known as single nucleotide
polymorphisms (SNPs).
Restriction enzymes are also used to digest genomic
DNA for gene analysis by Southern blot. This
technique allows researchers to identify how many
copies of a gene are present in the genome of one
individual, or how many gene mutations have
occurred within a population. This is called
restriction fragment length polymorphism
Activity of restriction enzyme
Mallikarjun School

Biology project
Topic:- Restriction enzyme

Submitted to:
Mr Manavendra Goswami

Submitted by:
Arnav Bhatt
Class XII Biology
Roll.no 6
Content
1.Restriction enzyme
2. History
3. Recognitation site
4. Types
5. Nomenclature
6. Working
7.Applications
8. Activity of restriction enzyme

Reference
 Class XII biology NCERT
 www.wikipedia.com
 www.khanacademy.com
 plantcellbiology.masters.grkraj.org
 www.britannica.com
 Premierbiosoft.com
 www.bio.miami.edu