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Molecular Scissors
Presented By:-
Avni Jethva
M.Sc Microbiology
(Sem2)
Content
• Introduction
• History
• Restriction enzymes
• Nomenclature
• Different recognition sequences
• Classification
• Applications
Introduction
Bacteria possess restriction systems as
protection mechanism against incoming
DNA.
• This system employs restriction enzymes
like endo- & exo- nucleases.
• Nucleases cleave phosphodiester bonds
between nucleotides.
History
• 1962-Arber & Dussoix :
E.coli can restrict lambda
phage DNA
• 1965-Arber: E.coli
methylate their DNA
• 1968-1st restriction
endonuclease in E.coli
discovered
• 1970-Smith, Wilcox &
Kelly: Discovered 1st DNA-
specific restriction
endonuclease in
H.influenzae
Restriction enzymes
• Specific endonucleases
• Recognize specific short sequences of DNA and
cleave the DNA at or near the recognition
sequence
• Recognition sequences: usually 4 or 6 bases but
there are some that are 5, 8, or longer
• Recognition sequences are palindromes
• Palindrome: sequence of DNA that is the same
when one strand is read from left to right or the
other strand is read from right to left– consists of
adjacent inverted repeats
Blunt & Sticky ends
Nomenclature
• Restriction enzymes are named based on the bacteria
in which they are isolated in the following manner:
• The EcoRV
endonuclease
has two sub-
units (shown in
blue and gray).
• The DNA is
highlydistorted
…
Applications
• rDNA Technology
• Plasmid Insertion
• Southern Blotting
• DNA Sequencing
• PCR
DNA Sequencing.
PCR