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Related Experiment Videos

The nucleotide sequence recognized by the BstEII restriction endonuclease.

J A Lautenberger, M H Edgell, C A Hutchison

    Gene
    |December 1, 1980
    PubMed
    Summary

    The BstEII endonuclease recognizes a specific seven-letter DNA sequence. This enzyme creates DNA fragments with five-nucleotide-long cohesive ends, useful in molecular biology.

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    Area of Science:

    • Molecular Biology
    • Enzymology
    • Genetics

    Background:

    • Restriction endonucleases are enzymes that cleave DNA at specific recognition sites.
    • BstEII is a type IIs restriction enzyme known for its unique recognition sequence and cleavage pattern.

    Purpose of the Study:

    • To characterize the precise restriction site recognized by the BstEII endonuclease.
    • To define the nature of the cohesive termini generated by BstEII digestion.

    Main Methods:

    • Bioinformatic analysis of known BstEII restriction sites.
    • Experimental validation of the proposed recognition sequence.
    • Analysis of DNA fragments generated by BstEII digestion to determine termini length and type.

    Main Results:

    • The BstEII restriction site is a heptameric sequence (5'-GTYRAC-3').
    • BstEII generates five-nucleotide-long cohesive termini upon cleavage.
    • The cleavage occurs 1 and 5 nucleotides downstream from the 5' and 3' ends of the recognition site, respectively.

    Conclusions:

    • The precise BstEII recognition sequence and cleavage mechanism have been elucidated.
    • Understanding the BstEII restriction site and its cohesive ends is crucial for its application in molecular cloning and genome engineering.

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