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Biospecific fractionation matrices for sequence specific endonucleases.

J George, J G Chirikjian

    Nucleic Acids Research
    |July 1, 1978
    PubMed
    Summary
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    Two novel affinity matrices, pyran and Cibacron Blue F3GA, effectively fractionate type II restriction endonucleases. These reusable matrices offer rapid purification of enzymes from crude extracts.

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Enzymology

    Background:

    • Restriction endonucleases are crucial tools in molecular biology.
    • Efficient purification methods are needed for these enzymes.
    • Existing methods can be time-consuming and costly.

    Purpose of the Study:

    • To develop novel biospecific affinity matrices for restriction endonuclease purification.
    • To evaluate the efficacy of pyran and Cibacron Blue F3GA matrices.
    • To assess the matrices' capacity, reusability, and development time.

    Main Methods:

    • Fractionation of type II restriction endonucleases using pyran and Cibacron Blue F3GA matrices.
    • Matrices synthesized by insolubilizing compounds to sepharose via cyanogen bromide linkage.

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  • Application of matrices for enzyme purification from crude extracts post-nucleic acid removal.

    • Main Results:

    • Successful fractionation of several type II restriction endonucleases was achieved.
    • The affinity matrices demonstrated high capacity in subsequent purification steps.
    • Matrices showed rapid development time and reusability, reducing purification costs.

    Conclusions:

    • Pyran and Cibacron Blue F3GA are effective biospecific matrices for restriction endonuclease purification.
    • These matrices offer an efficient, cost-effective, and reusable alternative for enzyme isolation.
    • The developed method simplifies the purification of essential molecular biology enzymes.