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Molecular cloning of seal myoglobin mRNA

D Wood, A Blanchetot, A J Jeffreys

    Nucleic Acids Research
    |November 25, 1982
    PubMed

    Insights

    Researchers isolated messenger RNA (mRNA) from grey seal muscle to study myoglobin. They found that seal myoglobin is encoded by a single gene, producing a 1400-nucleotide mRNA molecule.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Myoglobin is a protein found in skeletal muscle, crucial for oxygen storage.
    • Understanding gene expression and mRNA structure provides insights into protein synthesis and regulation.

    Purpose of the Study:

    • To isolate and characterize complementary DNA (cDNA) encoding grey seal myoglobin.
    • To investigate the gene structure and mRNA size of seal myoglobin.

    Main Methods:

    • Extraction of poly(A)+ RNA from grey seal skeletal muscle.
    • In vitro translation to identify myoglobin production.
    • Complementary DNA (cDNA) synthesis via reverse transcription and cloning into plasmid pAT 153.
    • DNA sequencing of cDNA inserts and hybridization experiments.

    Main Results:

    • Successfully generated cDNA clones containing myoglobin inserts from grey seal muscle RNA.
    • Sequence analysis revealed an incomplete cDNA clone (pSM 178) with 293 nucleotides of 3' non-translated mRNA sequences.
    • Hybridization experiments indicated that seal myoglobin is encoded by a single gene.
    • The mRNA for seal myoglobin is approximately 1400 nucleotides long, which is larger than related hemoglobin mRNAs.

    Conclusions:

    • The study successfully cloned and partially sequenced cDNA for grey seal myoglobin.
    • Seal myoglobin is transcribed from a single gene into a relatively large mRNA molecule.

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