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Types of Restriction Enzymes
Presentation · December 2020
DOI: 10.13140/RG.2.2.18102.52809
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Nidhi Puranik
Bharathiar University
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BARKATULLAH UNIVERSITY
Department of Biochemistry & Genetics
M.Sc - Molecular Biology
Paper code: M303: Genetic Engineering
Unit: I
Topic: DNA: Types of Restriction Enzymes
BY
Ms. NIDHI PURANIK , PhD
Faculty Member
Department of Biochemistry & Genetics
Lecture 4: 23.11.2020
Types of Restriction Endonucleases
There are 3 main categories of restriction Endonuclease enzymes:
1. Type-I
2. Type-II
3. Type-III
Type-I Restriction Endonucleases:
These are the complex type of endonucleases which cleave only one strand of DNA.
These enzymes have the recognition sequences of about 15 bp length. They require
Mg++ ions and ATP for their functioning. Such types of restriction endonucleases cleave
the DNA about 1000 bp away from the 5′-end of the sequence TCA located within the
recognition site. Important examples of Type-I restriction endonuclease enzymes are
EcoK, EcoB, etc.
Lecture 4: 23.11.2020
Type-II Restriction Endonucleases:
• These are most important endonucleases for gene cloning and hence for Recombinant
DNA Technology.
• These enzymes are most stable.
• They show cleavage only at specific sites and therefore they produce the DNA fragments
of a defined length.
• These enzymes show cleavage in both the strands of DNA, immediately outside the
recognition sequences.
• They require Mg++ ions for their functioning.
• Such enzymes are advantageous because they do not require ATP for cleavage and they
cause cleavage in both strands of DNA.
• Only Type II Restriction Endonucleases are
Lecture used for gene cloning due to their suitability.
4: 23.11.2020
• The recognition sequences for Type-II Restriction Endonuclease enzymes are in the
form of palindromic sequences with rotational symmetry, i.e., the base sequence in the
first half of one strand of DNA is the mirror image of the second half of other strand of
that DNA double helix.
• Important examples of Type-II Restriction endonucleases include Hinfl, EcoRI,
PvuII, Alul, Haelll, etc.
Lecture 4: 23.11.2020
Type-III Restriction Endonucleases
• These restriction enzymes are not used for gene cloning.
• They are the intermediate enzymes between Type-I and Type-II restriction
endonuclease.
• They require Mg++ ions and ATP for cleavage and they cleave the DNA at well-defined
sites in the immediate vicinity of recognition sequences, e.g. Hinf III, etc.
Lecture 4: 23.11.2020
Nomenclature of restriction enzymes
• Restriction enzymes are named based on the organism in which they were discovered.
• For example, the enzyme Hind III was isolated from the bacterium Haemophilus
influenzae, strain Rd.
• The first three letters (Hin) of the name are italicized because they abbreviate the genus
(H) and species (in) names of the organism. The fourth letter (d) typically comes from
the bacterial strain designation.
• Typically, the Roman numerals (I, II, III, etc.) indicates the order in which restriction
enzymes were discovered in a particular strain.
Lecture 4: 23.11.2020
Nomenclature of restriction enzymes consists three parts:
It indicates the genus and species of specific organism by using abbreviation into
three letters.
It indicates the strain of the relevant species by using a letter, number or its
combination.
It indicates specific restriction modification systems present in the same organism or
strain by using a roman numeral.
Example: the name of EcoRI restriction enzyme is derived as: Abbreviation
Corresponding full form
E Escherichia (genus)
Co coli (species)
R RY13 (strain)
I First identified in the bacterium
Lecture 4: 23.11.2020
Lecture 4: 23.11.2020
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