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Related Experiment Videos

[Site-specific endonucleases LplI and AagI].

N N Sokolov, Z P Manialene, V V Butkus

    Bioorganicheskaia Khimiia
    |August 1, 1990
    PubMed
    Summary

    Two novel restriction endonucleases, LplI and AagI, were identified from Lactobacillus plantarum and Achromobacter agile. These enzymes are isoschizomers of ClaI, recognizing the ATCGAT sequence.

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    Thermostable Recombinant Polypeptides as the Source of L-Amino Acids for Culture Media.

    Bulletin of experimental biology and medicine·2018

    Area of Science:

    • Molecular Biology
    • Enzymology
    • Microbial Genetics

    Context:

    • Restriction endonucleases are crucial tools in molecular biology for DNA manipulation.
    • The discovery of new endonucleases expands the repertoire available for genetic engineering and research.
    • Characterizing novel enzymes is essential for understanding DNA recognition and cleavage mechanisms.

    Purpose:

    • To isolate and characterize novel site-specific endonucleases from microbial sources.
    • To determine the DNA recognition and cleavage sites of the newly identified enzymes.
    • To assess the potential of Lactobacillus plantarum for large-scale production of the LplI enzyme.

    Summary:

    • New site-specific endonucleases, LplI and AagI, were purified from Lactobacillus plantarum and Achromobacter agile.

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  • Both enzymes recognize and cleave the ATCGAT sequence, identifying them as isoschizomers of the known ClaI restriction endonuclease.
  • Purification involved polyethylenimine treatment, phase fractionation, and multiple chromatography steps.
  • The Lactobacillus plantarum strain exhibits significantly higher endonuclease productivity compared to the ClaI producer, indicating its potential for preparative LplI isolation.

    • Impact:

    • The identification of LplI and AagI expands the available tools for restriction digestion, particularly for the ATCGAT sequence.
    • L. plantarum emerges as a promising host for the efficient, large-scale production of the LplI endonuclease.
    • This discovery facilitates advancements in DNA analysis, genetic engineering, and synthetic biology applications.