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    Restriction Enzymes Practice

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    Oktira Roka Aji
    Restriction enzymes are proteins produced by bacteria that cut DNA at specific short nucleotide sequences called restriction sites. Each restriction enzyme recognizes a different restriction site. Cutting DNA at these sites allows bacteria to defend against invading viruses and other bacteria. Restriction enzymes leave different types of ends - "sticky ends" that can rejoin with other cut DNA pieces, or "blunt ends" that cannot. These enzymes are useful for DNA fingerprinting and cutting and rejoining DNA in genetic engineering.

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    Restriction Enzymes Practice

    Uploaded by

    Oktira Roka Aji
    120 views2 pages
    Restriction enzymes are proteins produced by bacteria that cut DNA at specific short nucleotide sequences called restriction sites. Each restriction enzyme recognizes a different restriction site. Cutting DNA at these sites allows bacteria to defend against invading viruses and other bacteria. Restriction enzymes leave different types of ends - "sticky ends" that can rejoin with other cut DNA pieces, or "blunt ends" that cannot. These enzymes are useful for DNA fingerprinting and cutting and rejoining DNA in genetic engineering.

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    Restriction enzyme

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    Restriction enzymes are proteins produced by bacteria that cut DNA at specific short nucleotide sequences called restriction sites. Each restriction enzyme recognizes a different restriction site. Cutting DNA at these sites allows bacteria to defend against invading viruses and other bacteria. Restriction enzymes leave different types of ends - "sticky ends" that can rejoin with other cut DNA pieces, or "blunt ends" that cannot. These enzymes are useful for DNA fingerprinting and cutting and rejoining DNA in genetic engineering.

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    © All Rights Reserved
    Download as DOC, PDF, TXT or read online from Scribd
    Download as doc, pdf, or txt
    120 views2 pages

    Restriction Enzymes Practice

    Uploaded by

    Oktira Roka Aji
    Restriction enzymes are proteins produced by bacteria that cut DNA at specific short nucleotide sequences called restriction sites. Each restriction enzyme recognizes a different restriction site. Cutting DNA at these sites allows bacteria to defend against invading viruses and other bacteria. Restriction enzymes leave different types of ends - "sticky ends" that can rejoin with other cut DNA pieces, or "blunt ends" that cannot. These enzymes are useful for DNA fingerprinting and cutting and rejoining DNA in genetic engineering.

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    Restriction Enzymes: DNA Scissors

    Background : DNA fingerprinting is made possible in part by special enzymes that cut DNA. These enzymes are called
    restriction enzymes. Restriction enzymes are proteins that bacteria use to cut up DNA that doesn’t belong to them. If a
    bacterium senses that a virus is trying to invade, or a different species of bacterium represents a threat, it can use a restriction
    enzyme to cut up the foreigner’s DNA.

    Restriction enzymes don’t just cut DNA randomly – that would lead to the destruction of the bacterium’s own DNA. (That’s a
    bad thing!) Restriction enzymes recognize specific segments of bases called restriction sites. Each different restriction
    enzyme (and there are hundreds, made by different species of bacteria) has its own particular restriction site where it will cut
    DNA. Most restriction sites are 4 to 6 bases long and are DNA palindromes. A DNA palindrome is a sequence where the “top”
    strand reads the same as the “bottom” strand running in the opposite direction. For example:

    This sequence happens to be the restriction site for the enzyme called EcoR1. Its name comes from the bacterium in which it
    was discovered – Escherichia coli strain RY 13 (that’s the EcoR part). The “1” comes from the fact that it was the first
    restriction enzyme found in this strain of E. coli. The enzyme EcoR1 cuts between the A and G in each strand of the DNA.
    (The arrows in the diagram below show where Eco R1 snips the DNA.) After the cuts are made, the two strands are
    held together only by weak hydrogen bonds between complementary bases, which separate easily, like a broken zipper.

    You may notice that the EcoR1 enzyme does not cut straight across the DNA molecule. When EcoR1 cuts DNA, it leaves
    single stranded “tails” called sticky ends, because they could stick (although not very tightly) to other segments of DNA that
    have been cut with the same enzyme. (Remember this trait of restriction enzymes, because these sticky ends can
    be useful for re-joining the cut DNA – even if the DNA pieces originally came from different species.) Not all
    restriction enzymes make sticky ends when they cut; some cut straight across the DNA to leave blunt ends that won’t stick to
    each other.

    The restriction sites for four other restriction enzymes are shown below. The arrows show where they cut the DNA strands:
    Restriction Enzymes
    Directions: Identify the restriction sites for each of the examples given. Show the cuts , sticky
    ends or blunt, number of DNA fragments produced and the number of base pairs in each
    (count the top row). (If there are three nucleotides on either side of the dash it is a blunt cut. If there are fewer
    than three bases on either side of the dash it produces “sticky ends” cut.)
    Remember: Analyze in the 5’ 3’ direction

    1. HindIII--- 5' GTC ↓GAC 3'

    5' ACGACGTAGTCGACTTATTAT GTCGACCCGCCGCGTGTCGACCATCA 3'


    3' TGCTGCATCAGCTGAATAATACAGCTGGGCGGCGCACAGCTGGTAGT 5'

    Number of pieces of DNA _______

    2. EcoRI --- 5' G ↓AATTC 3'

    5' ACG ACGTATTAGAATTCTTAT CCGCCGCCGGAATTCT CATCA 3'


    3' TGC TGCATAATCTTAAGAATAGGCGGCGGCCTTAAGAGTAGT 5'

    Number of pieces of DNA _______

    3. HaeIII --- 5' CC ↓ GG 3'

    5' ACGCCGGCCGTATTAT CCGGATCCGCCG CCGGCTGTCCCGGATCA 3'


    3' TGCGGCCGGCATAATAGGCCTAGGCGGCGGCCGACAGGGCCTAGT 5'

    Number of pieces of DNA _______

    4. BamI --- 5' CCTAG ↓G 3'

    5' ACGCCTAGGACGTATTATCCTAGGTAT CCGCCGCCGT CATCA 3'


    3' TGCGGATCCTGCATAATAGGATCCATAGGCGGCGGCAGTAGT 5'

    Number of pieces of DNA _______

    5. HindII --- 5' GTC ↓- GAC 3' and HaeIII --- 5' CC ↓ GG 3'

    5' ACGGTCGACACGTATTATTAGTCGACTCCGCCGCCGCCGGTCATCA 3'


    3' TGCCAGCTGTGCATAATAATCAGCTGAGGCGGCGGCGGCCAGTAGT 5'

    Number of pieces of DNA _______


    6. HindII --- 5' GTC ↓ GAC 3', HaeIII --- 5' CC ↓ GG 3' and BamI --- 5' CCTAG ↓ G 3'

    5' ACGCCGGACGTACCTAGGTTTAGTCGACTC CGCCG CCCCTAGGGTCATCA 3'


    3' TGCGGCCTGCATGGATCCAAATCAGCTGAGGCGGCGGGGATCCCAGTAGT 5'

    Number of pieces of DNA _______

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