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Related Experiment Videos

A new sequence-specific endonuclease (Bsp) from Bacillus sphaericus.

A Kiss, B Sain, E Csordás-Tòth

    Gene
    |July 1, 1977
    PubMed
    Summary

    Researchers isolated a new restriction enzyme, Bsp, from Bacillus sphaericus R. This enzyme cleaves DNA similarly to Bsu and HaeIII, offering a high-yield purification process.

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    Area of Science:

    • Molecular Biology
    • Enzymology
    • Biochemistry

    Background:

    • Restriction endonucleases are crucial tools in molecular biology for DNA manipulation.
    • Identifying novel enzymes with unique specificities expands the toolkit for genetic engineering and research.

    Purpose of the Study:

    • To isolate and characterize a new restriction endonuclease from Bacillus sphaericus R.
    • To determine the cleavage specificity and purification efficiency of the novel enzyme.

    Main Methods:

    • Enzyme isolation and purification using Bio-Gel filtration, ammonium sulfate fractionation, and phosphocellulose chromatography.
    • DNA digestion assays using phiX174 replicative form DNA.
    • Analysis of cleavage products to determine specificity and terminal nucleotides.

    Main Results:

    • A novel restriction endonuclease, designated Bsp, was purified from Bacillus sphaericus R.
    • The enzyme preparation was free of non-specific nucleases after phosphocellulose chromatography.
    • Bsp exhibits the same cleavage specificity as Bacillus subtilis (Bsu) and Haemophilus aegyptius (HaeIII) restriction endonucleases.
    • The 5'-terminal nucleotide of cleavage products is cytosine (C).
    • Bacillus sphaericus R produces Bsp in large quantities, facilitating high-yield purification.

    Conclusions:

    • The newly isolated Bsp restriction endonuclease is a valuable tool for DNA analysis and manipulation.
    • Its high yield and purification efficiency make it an attractive enzyme for biotechnological applications.
    • The characterized specificity expands the repertoire of restriction enzymes available to researchers.

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