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Related Concept Videos

Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
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Restriction Enzymes

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Updated: Jul 17, 2026

Screening Foodstuffs for Class 1 Integrons and Gene Cassettes
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Published on: June 19, 2015

Multiple steps in DNA recognition by restriction endonuclease from E. coli K.

R Yuan, T A Bickle, W Ebbers

    Nature
    |August 14, 1975
    PubMed
    Summary

    Restriction endonuclease from E. coli K binds DNA through three distinct complexes: initial, recognition, and cleavage. ATP is crucial for the final cleavage complex formation during DNA processing.

    Area of Science:

    • Molecular Biology
    • Enzymology
    • Genetics

    Background:

    • Restriction endonucleases are key enzymes in molecular biology, involved in DNA defense mechanisms.
    • Understanding DNA recognition by these enzymes is crucial for genetic engineering and biotechnology.
    • The restriction endonuclease from E. coli K (EcoK) is a well-studied example with complex DNA interactions.

    Purpose of the Study:

    • To elucidate the sequential mechanism of DNA recognition and binding by the activated restriction endonuclease from E. coli K.
    • To experimentally differentiate the distinct enzyme-DNA complexes involved in the EcoK restriction process.
    • To identify the role of ATP in the formation of the cleavage complex.

    Main Methods:

    • Biophysical techniques were employed to study enzyme-DNA interactions.

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  • Experimental differentiation of three distinct enzyme-DNA complex intermediates.
  • Investigation of ATP-dependent complex formation.

    • Main Results:

    • Three sequential enzyme-DNA complexes were identified: an initial nonspecific complex, a specific recognition complex at the host specificity site, and an ATP-dependent cleavage complex.
    • The formation of the cleavage complex is contingent upon the presence of ATP.
    • The study provides a stepwise model for EcoK DNA binding and cleavage.

    Conclusions:

    • The DNA recognition process by EcoK involves a series of distinct, experimentally verifiable complexes.
    • ATP plays a critical role in facilitating the transition from recognition to cleavage.
    • This detailed understanding of EcoK mechanism aids in the application of restriction enzymes in molecular biology.