Restriction Enzymes

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Concept Tutorial
Restriction Enzyme
 Restriction enzyme is an endonuclease which cut DNA at a particular
sequence.
 Because their nuclease activity is restricted to specific sequences, they are
called restriction enzymes.
 As they are able to make cut at desired location in DNA strand hence also
called “Molecular scissors”.
 Their recognition sequences are palindromic nucleotide sequences i.e.
group of letters that form the same words when read both forward and
backward e.g. “MADAM”.
Types of Cut Produced by Restriction Enzymes
Action of Restriction Enzyme
Exonuclease
vs.
Endonuclease
Convention for naming Restriction
Enzyme

 In EcoRI, “E” stands for Escherichia and “co” stands

for coli.
 The letter ‘R’ is derived from the name of strain
(Rough).
 Roman numbers indicate the order in which the
enzymes were isolated from that strain of bacteria.
 DNA
Cloning
 Restrictionendonuclease acts as molecular scissors and cuts the DNA at specific sites to
separate desired gene.
 Segmentof desired gene is joined to plasmid with the help of DNA ligase. This composite
DNA is called Recombinant DNA.
 Introducing recombinant DNA into host cells.
 Selecting host cells that contain recombinant DNA.
 Maintaining recombinant host cells in sterile culture medium.
Note:
 Themethods used to accomplish these tasks are collectively known as Recombinant DNA
technology or Genetic engineering.
 Escherichia
coli was the first organism used for recombinant DNA work and still is the most
common host cell.