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Characterization of fatty acid synthetase cDNA clone and its mRNA

C Yan, E A Wood, J W Porter

    Biochemical and Biophysical Research Communications
    |February 15, 1985
    PubMed

    Insights

    Researchers identified four complementary DNA (cDNA) clones for fatty acid synthetase (FAS) mRNA. Analysis revealed a single gene copy for FAS, with distinct mRNA transcripts detected in both cytoplasmic and nuclear RNA.

    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Genetics

    Background:

    • Fatty acid synthetase (FAS) is a crucial enzyme in fatty acid biosynthesis.
    • Understanding the genetic and transcriptomic landscape of FAS is vital for metabolic research.

    Purpose of the Study:

    • To identify and characterize cDNA clones encoding fatty acid synthetase (FAS) mRNA.
    • To analyze the transcript size and gene copy number of FAS.

    Main Methods:

    • Hybrid-select translation was used to identify cDNA clones complementary to FAS mRNA.
    • Restriction mapping was performed on identified cDNA clones.
    • Northern blot analysis was conducted on cytoplasmic and nuclear RNA.
    • Southern blot analysis was performed on genomic DNA.

    Main Results:

    • Four cDNA clones (pFAS-7, pFAS-17, pFAS-18, and pFAS-15) were identified, with three sharing common sequences.
    • Northern analysis revealed two major mRNA transcript sizes (9.2 Kb and 8.4 Kb) in cytoplasmic RNA and potential unprocessed transcripts (14 Kb and 11 Kb) in nuclear RNA.
    • Southern analysis indicated a single gene copy for fatty acid synthetase in the genome.

    Conclusions:

    • The identified cDNA clones represent different regions of the fatty acid synthetase mRNA.
    • The presence of multiple transcript sizes suggests potential alternative splicing or processing events.
    • The fatty acid synthetase gene is present as a single copy, providing a basis for further functional studies.

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