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Cloning muscle isoforms of neural cell adhesion molecule using an episomal shuttle vector

Somatic Cell and Molecular Genetics volume 18, pages 163–177 (1992)Cite this article

Abstract

Three distinct transcripts encoding two phosphatidylinositol (PI) linked isoforms of the neural cell adhesion molecule (NCAM) are induced during the differentiation of C2C12 myoblasts into myotubes. Corresponding NCAM clones were isolated from a mouse muscle cDNA library made in an Epstein-Barr virus shuttle vector that replicates extrachromosomally in human cells. Following transfection with the library, human cells expressing mouse NCAM were enriched using the fluorescence-activated cell sorter. Episomal NCAM clones recovered from sorted cells contain an 18-bp insert between exons 12 and 13. Two other NCAM cDNAs encode identical polypeptides containing a 108-bp insert homologous to the complete MSD1 domain, but differ in their 3′ untranslated regions. Induction of MSD1-containing transcripts in advance of myotube formation suggests that muscle-specific NCAMs contribute to myogenesis from the earliest stages of differentiation. Moreover, our studies demonstrate the feasibility of cloning tissue-specific molecules by transfection and expression of cDNA libraries in episomal vectors.

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Author information

Author notes

  1. Lydia C. Pan

    Present address: Research Division, The Hospital for Special Surgery, 535 East 70th Street, 10021, New York, New York

Affiliations

  1. Pharmacology Department, Stanford University School of Medicine, 94305, Stanford, California

    Lydia C. Pan & Helen M. Blau

  2. Department of Neurosciences, Roche Institute of Molecular Biology, Roche Research Center, 07110, Nutley, New Jersey

    Robert F. Margolskee

Authors
  1. Lydia C. Pan
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  2. Robert F. Margolskee
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  3. Helen M. Blau
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Pan, L.C., Margolskee, R.F. & Blau, H.M. Cloning muscle isoforms of neural cell adhesion molecule using an episomal shuttle vector. Somat Cell Mol Genet 18, 163–177 (1992). https://doi.org/10.1007/BF01233162

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  • DOI: https://doi.org/10.1007/BF01233162

Keywords

  • Polypeptide
  • Human Cell
  • cDNA Library
  • Untranslated Region
  • Cell Adhesion Molecule