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    Restriction Enzymes

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    skmfsc
    Restriction enzymes recognize specific short DNA sequences called restriction sites and cleave DNA molecules at these sites. The enzyme BamHI recognizes the palindrome sequence 5'-GGATCC-3' and cuts within this sequence, leaving "sticky ends". Restriction enzymes can cut DNA asymmetrically, leaving overhangs, or symmetrically at the same position on each strand. They cut DNA from any source as long as the restriction site is present. This cleavage produces restriction fragments that can be analyzed by gel electrophoresis.

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    Restriction Enzymes

    Uploaded by

    skmfsc
    164 views9 pages
    Restriction enzymes recognize specific short DNA sequences called restriction sites and cleave DNA molecules at these sites. The enzyme BamHI recognizes the palindrome sequence 5'-GGATCC-3' and cuts within this sequence, leaving "sticky ends". Restriction enzymes can cut DNA asymmetrically, leaving overhangs, or symmetrically at the same position on each strand. They cut DNA from any source as long as the restriction site is present. This cleavage produces restriction fragments that can be analyzed by gel electrophoresis.

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    restriction enzymes and specific DNA cleavage

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    Restriction enzymes

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    Restriction enzymes recognize specific short DNA sequences called restriction sites and cleave DNA molecules at these sites. The enzyme BamHI recognizes the palindrome sequence 5'-GGATCC-3' and cuts within this sequence, leaving "sticky ends". Restriction enzymes can cut DNA asymmetrically, leaving overhangs, or symmetrically at the same position on each strand. They cut DNA from any source as long as the restriction site is present. This cleavage produces restriction fragments that can be analyzed by gel electrophoresis.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    164 views9 pages

    Restriction Enzymes

    Uploaded by

    skmfsc
    Restriction enzymes recognize specific short DNA sequences called restriction sites and cleave DNA molecules at these sites. The enzyme BamHI recognizes the palindrome sequence 5'-GGATCC-3' and cuts within this sequence, leaving "sticky ends". Restriction enzymes can cut DNA asymmetrically, leaving overhangs, or symmetrically at the same position on each strand. They cut DNA from any source as long as the restriction site is present. This cleavage produces restriction fragments that can be analyzed by gel electrophoresis.

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    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
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    Restriction enzymes recognize specific palindromic sequences and cleave DNA at those sites.

    The recognition sequence for BamHI is 5'-GGATCC-3'.

    Restriction enzymes can cleave DNA asymmetrically or symmetrically, leaving sticky or blunt ends.

    Restriction Enzymes and SiteSpecific DNA Cleavage

    a class of enzymes known as restriction endonucleases or


    restriction enzymes are able to cleave DNA molecules at the
    positions at which particular, short sequences of bases are
    present.
    For example, the enzyme BamHI recognizes the double-stranded
    sequence
    5'-GGATCC-3'
    3'-CCTAGG-5'

    Mechanism of DNA cleavage by the restriction enzyme BamHI. The enzyme


    makes a single cut in the backbone of each DNA strand wherever the duplex
    contains a BamHI restriction site

    The nucleotide sequence recognized for cleavage by a restriction


    enzyme is called the restriction site of the enzyme

    The restriction enzymes cleave their restriction site asymmetrically


    (at different sites in the two DNA strands), but some restriction
    enzymes cleave symmetrically (at the same site in both strands).
    The former leave sticky ends because each end of the cleaved site
    has a small, single-stranded overhang that is complementary in base
    sequence to the other end.

    In virtually all cases, the restriction site of a restriction enzyme


    reads the same on both strands, provided that the opposite polarity
    of the strands is taken into account; for example, each strand in the
    restriction site of BamHI reads 5'-GGATCC-3.
    A DNA sequence with this type of symmetry is called a
    palindrome. (In ordinary English, a palindrome is a word or
    phrase that reads the same forwards and backwards, such as
    ''madam.")

    Table -Some restriction endonucleases, their sources, and their cleavage sites

    Note: The vertical dashed line indicates the axis of symmetry in each sequence. Red
    arrows indicate the sites of cutting. The enzyme TaqI yields cohesive ends consisting of
    two nucleotides, whereas the cohesive ends produced by the other enzymes contain four
    nucleotides. Pu and Py refer to any purine and pyrimidine, respectively.

    Restriction enzymes have the following important


    characteristics:
    Most restriction enzymes recognize a single restriction site.
    The restriction site is recognized without regard to the source of
    the DNA.
    Because most restriction enzymes recognize a unique restriction
    site sequence, the number of cuts in the DNA
    from a particular organism is determined by the number of
    restriction sites present.

    The DNA fragment produced by a pair of adjacent cuts in a


    DNA molecule is called a restriction fragment. A large
    DNA molecule will typically be cut into many restriction
    fragments of different sizes.
    For example, an E. coli DNA molecule, which contains 4.7
    106 base pairs, is cut into several hundred to several
    thousand fragments, and mammalian nuclear DNA is cut
    into more than a million fragments

    A map showing the unique sites of cutting of the DNA of a


    particular organism by a single enzyme is called a restriction
    map.
    The family of fragments produced by a single enzyme can be
    detected easily by gel electrophoresis, and particular DNA
    fragments can be isolated by cutting out the small region of the
    gel that contains the fragment and removing the DNA from the
    gel.

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