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A second site-specific restriction endonuclease from Staphylococcus aureus.

J S Sussenbach, P H Steenbergh, J A Rost

    Nucleic Acids Research
    |April 1, 1978
    PubMed
    Summary
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    A novel restriction enzyme, Sau96 I, was isolated from Staphylococcus aureus. This enzyme targets a specific DNA sequence (5'--G-G-N-C-C--3') and shows activity against various viral DNAs, suggesting a role in restriction-modification.

    Area of Science:

    • Molecular Biology
    • Enzymology
    • Genetics

    Background:

    • Restriction endonucleases are crucial tools in molecular biology for DNA manipulation.
    • Understanding new restriction enzymes aids in expanding the repertoire for genetic engineering and analysis.
    • Staphylococcus aureus is a known source of various bioactive compounds, including enzymes.

    Purpose of the Study:

    • To isolate and characterize a novel site-specific restriction endonuclease from Staphylococcus aureus PS 96.
    • To determine the DNA recognition sequence and cleavage pattern of the isolated enzyme.
    • To investigate the potential role of the enzyme in biological restriction-modification systems.

    Main Methods:

    • Isolation and purification of the restriction endonuclease from Staphylococcus aureus PS 96.

    Related Experiment Videos

  • DNA cleavage assays using various viral DNA substrates (adenovirus type 5, lambda DNA, SV40 DNA, φX174 RF DNA).
  • Determination of the specific DNA recognition sequence (5'--G-G-N-C-C--3') and cleavage sites.

    • Main Results:

    • Successful isolation of a site-specific restriction endonuclease, designated Sau96 I.
    • Sau96 I recognizes and cleaves the degenerate sequence 5'--G-G-N-C-C--3'.
    • The enzyme exhibited multiple cleavage events on adenovirus type 5, lambda DNA, SV40 DNA, and φX174 RF DNA.

    Conclusions:

    • Sau96 I is a novel restriction enzyme with a unique recognition site.
    • The enzyme's activity on diverse viral DNAs supports its potential function in bacterial defense mechanisms.
    • Further studies are warranted to elucidate the precise role of Sau96 I in Staphylococcus aureus restriction-modification.