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Identification and DNA sequence of a human apolipoprotein E cDNA clone.

J L Breslow, J McPherson, A L Nussbaum

    The Journal of Biological Chemistry
    |December 25, 1982
    PubMed
    Summary

    Researchers identified human apolipoprotein E (apo-E) cDNA clones from liver tissue. DNA sequencing revealed single base substitutions explain apo-E genetic variations, supporting a mutation hypothesis.

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Apolipoprotein E (apo-E) plays a crucial role in lipid metabolism and is associated with various diseases.
    • Genetic variations in the apo-E gene lead to different apo-E isoforms, influencing lipoprotein metabolism.
    • Understanding the molecular basis of apo-E polymorphism is essential for disease risk assessment.

    Purpose of the Study:

    • To isolate and characterize cDNA clones encoding human apolipoprotein E.
    • To investigate the genetic basis of apolipoprotein E polymorphism at the DNA sequence level.

    Main Methods:

    • Screening of an adult human liver cDNA library using a degenerate oligonucleotide probe.
    • PstI digestion and DNA sequencing of selected cDNA clones.
    • Analysis of apo-E polymorphic sites at amino acids 112, 145, and 158.

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    Main Results:

    • Identification and characterization of cDNA clones encoding human apo-E, with inserts of approximately 900 base pairs.
    • DNA sequence analysis confirmed the presence of polymorphic sites and revealed single base substitutions within the apo-E gene.
    • The observed apo-E phenotypes can be explained by specific single base substitutions in the codons for amino acids 112, 145, and 158.

    Conclusions:

    • The study successfully identified human apo-E cDNA clones and elucidated the molecular basis of apo-E polymorphism.
    • The findings support the hypothesis that apo-E genetic variations arise from mutations within the apo-E structural gene.
    • This research provides a foundation for understanding the genetic determinants of apo-E function and its role in health and disease.