Abdul Wali Khan University
Mardan, Pakistan
Submitted by:
Waseem Feroz
Roll no 09
Semester 5th
Department of Microbiology
Subject: Biotechnology
Submitted To:
Sir, Dr. Zamarud Shah
Assignment:Restrictions enzymes.
Restriction Enzymes:
Restriction enzymes are proteins produced by bacteria that
cleave DNA at specific recognition sequences known as
restriction sites. These enzymes play a crucial role in molecular
biology, particularly in genetic engineering and cloning.
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Naming Convention
The names of restriction enzymes typically follow a specific
convention:
• First two letters: Abbreviation of the bacterial genus from
which the enzyme was isolated.
• Third letter: Abbreviation of the bacterial species.
• Roman numeral: Indicates the order of discovery of the
enzyme from that organism.
For example, EcoRI is derived from Escherichia coli (strain
RY13), the first restriction enzyme discovered from this organism.
Sources
Restriction enzymes are primarily isolated from bacteria. Different
species of bacteria produce enzymes that recognize and cleave
different restriction sites. Some common sources of restriction
enzymes include:
• Escherichia coli
• Haemophilus influenzae
• HindIII
• BamHI
• EcoRV
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Recognition Sequences
Restriction enzymes recognize specific DNA sequences, typically
4-8 base pairs long, known as restriction sites. These sites are
usually palindromic, meaning they read the same backward and
forward on the opposite strand.
Example:
• EcoRI: 5' GAATTC 3' 3' CTTAAG 5'
When a restriction enzyme encounters its recognition site, it cuts
the DNA at a specific point within or near the site. This cleavage
can result in either blunt ends or sticky ends, depending on the
enzyme.
Blunt Ends: Both strands of DNA are cut at the same position,
resulting in a flat end. Sticky Ends: The DNA is cut in a
staggered manner, leaving single-stranded overhangs that can
base-pair with complementary sequences.
Applications of Restriction Enzymes
Restriction enzymes are widely used in molecular biology for
various applications, including:
• Cloning: Creating recombinant DNA molecules by joining
DNA fragments from different sources.
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• DNA fingerprinting: Identifying individuals based on
unique DNA fragment patterns.
• Gene analysis: Studying the structure and function of
genes.
• Genome sequencing: Determining the complete
nucleotide sequence of an organism's genome.
Enzyme Source Organism. Recognition
Sequence
EcoRI. Escherichia coli 5’
GAATTC 3’
HindIII. Haemophilus influenzae 5’
AAGCTT 3’
BamHI Bacillus amyloliquefaciens. 5’
GGATCC 3’
SalI Streptomyces albus 5’
GTCGAC 3’
XbaI. Xanthomonas albilineans. 5’ TCTAGA
3’
PstI Providencia stuartii 5’
CTGCAG 3’
SmaI. Serratia marcescens. 5’
CCCGGG 3
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