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Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction
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Intervening sequence of DNA identified in the structural portion of a mouse beta-globin gene

S M Tilghman, D C Tiemeier, J G Seidman

    Proceedings of the National Academy of Sciences of the United States of America
    |February 1, 1978
    PubMed

    Insights

    The mouse beta-globin gene contains DNA sequences, called intervening sequences, that interrupt its structure. These sequences explain the gene’s unusual appearance and suggest it is encoded in discontinuous segments.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • The structure of eukaryotic genes often differs from their messenger RNA (mRNA) transcripts.
    • Understanding gene organization is crucial for deciphering gene expression and regulation.

    Purpose of the Study:

    • To investigate the structural basis for the unusual appearance of a hybrid formed between mouse beta-globin mRNA and its cloned gene.
    • To identify and characterize intervening sequences within the mouse beta-globin gene.

    Main Methods:

    • Electron microscopy of a hybrid molecule formed between 9S mouse beta-globin mRNA and a cloned gene segment.
    • Analysis of restriction enzyme digestion patterns.
    • Nucleotide sequencing of the gene region spanning structural and intervening sequences.

    Main Results:

    • Electron microscopy revealed structural anomalies in the hybrid molecule.
    • The anomalies were attributed to at least one, and possibly two, intervening DNA sequences within the mouse beta-globin gene.
    • A large intervening sequence (~550 base pairs) was identified within the structural globin sequence after codon 104.
    • A smaller, putative intervening sequence was located near the 5' end of the coding sequence.

    Conclusions:

    • The mouse beta-globin gene is not a continuous coding sequence.
    • The gene is interrupted by intervening sequences, leading to a discontinuous structure.
    • The beta-globin gene appears to be encoded in two, or possibly three, discontinuous segments.

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