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Restriction Enzymes: Restriction Enzymes Is An Enzyme That Cuts Double Stranded or Single

Restriction enzymes are enzymes that cut DNA at specific recognition sequences. They recognize and cut DNA at particular sites, generating DNA fragments with either blunt or sticky ends. Sticky ends have short, single-stranded overhangs that allow fragments to recombine, while blunt ends have no overhangs. Restriction enzymes play an important role in DNA manipulation by cutting DNA into pieces that can be reassembled in new combinations.

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110 views3 pages

Restriction Enzymes: Restriction Enzymes Is An Enzyme That Cuts Double Stranded or Single

Restriction enzymes are enzymes that cut DNA at specific recognition sequences. They recognize and cut DNA at particular sites, generating DNA fragments with either blunt or sticky ends. Sticky ends have short, single-stranded overhangs that allow fragments to recombine, while blunt ends have no overhangs. Restriction enzymes play an important role in DNA manipulation by cutting DNA into pieces that can be reassembled in new combinations.

Uploaded by

Arafat Miah
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Restriction enzymes: Restriction enzymes is an enzyme that cuts double stranded or single

stranded DNA at a specific recognition nucleotide sequence (mostly palindromic or symmetry)


known as restriction site. It is also called Restriction endonuclease or Molecular scissor or
Bimolecular scissors.
Function of Restriction enzyme:
 Recognition of the signal sequence or restriction signal
 Cleave at the specific DNA segment
 Modify the self DNA molecule
 Restriction enzymes protect cells from foreign DNA
Properties of restriction fragmentation:
 Specificity
 Cleave at the specific DNA segment
 Can generate either sticky or blunt end
Mechanism of restriction fragmentations: Restriction enzymes recognize a specific sequence of
nucleotides, and produce two types of double-stranded cut in the DNA, these may be either Blunt
end or Sticky end.
Sticky end: Fragment end of a DNA molecule with a short single stranded overhang. Cut in the
site of the cleavage site. These end is produced followed by designed fashion. Sticky end is
produced by the following enzyme
EcoRI
HindIII
BamHI
MspI etc.
Blunt end: Fragment end of a DNA molecule with no overhang. Cut in the midde of the cleavage
site. These end is produced followed by direct fashion. Blunt end is produced bythe following
enzyme
AluI
HaeIII etc.
Given the DNA shown below …
5’ ATTGAGGATCCGGAATGTGTCCTGAATTCGCTCCACG 3’
3’ TAACTCCTAGGCCTTACACAGGACTTAAGCGAGGTGC 5’
If this DNA was cut with MspI, how many DNA fragments would you expect?
Answer:
Given below the cleavage site of MspI restriction enzyme

If the above DNA sequence was cut with MspI, I would except 4 DNA fragments (two double
stranded DNA fragments) and produce sticky end type fragments, these fragments are given
below:

5’ ATTGAGGATCCGGAATGTGTCCTGAATTCGCTCCACG 3’
3’ TAACTCCTAGGCCTTACACAGGACTTAAGCGAGGTGC 5’

5’ ATTGAGGATC 3’ 5’ CGGAATGTGTCCTGAATTCGCTCCACG 3’
3’ TAACTCCTAGGC 5’ 3’ CTTACACAGGACTTAAGCGAGGTGC 5’

Sticky end is produced by MspI

Noted down the sequence of these double stranded DNA fragments:


One double stranded DNA fragments

5’ ATTGAGGATC 3’
3’ TAACTCCTAGGC 5’

Another double stranded DNA fragments

5’ CGGAATGTGTCCTGAATTCGCTCCACG 3’
3’ CTTACACAGGACTTAAGCGAGGTGC 5’
Given the DNA shown below …
5’ ATTGAGGATCCGGAATGTGTCCTGAATTCGCTCCACG 3’
3’ TAACTCCTAGGCCTTACACAGGACTTAAGCGAGGTGC 5’
If this DNA was cut with EcoRI, how many DNA fragments would you expect?
Answer: Given below the cleavage site of EcoRI restriction enzyme

If the above DNA sequence was cut with EcoRI, I would except 4 DNA fragments (two double
stranded DNA fragments) and produce sticky end type fragments, these fragments are given
below:

5’ ATTGAGGATCCGGAATGTGTCCTGAATTCGCTCCACG 3’
3’ TAACTCCTAGGCCTTACACAGGACTTAAGCGAGGTGC 5’

5’ ATTGAGGATCCGGAATGTGTCCTG 3’ 5’ AATTCGCTCCACG 3’
3’ TAACTCCTAGGCCTTACACAGGACTTAA 5’ 3’ GCGAGGTGC 5’

Sticky end is produced by EcoRI

Noted down the sequence of these double stranded DNA fragments:


One double stranded DNA fragments

5’ ATTGAGGATCCGGAATGTGTCCTG 3’
3’ TAACTCCTAGGCCTTACACAGGACTTAA 5’

Another double stranded DNA fragments

3’ GCGAGGTGC 5’
5’ AATTCGCTCCACG 3’

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