Novel cDNA clones obtained by antibody screening of a mouse cerebellar cDNA expression library

M. Kambouris, L. C. Triarhou, S. R. Dlouhy, L. Sangameswaran, F. Luo, B. Ghetti, M. E. Hodes

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

In order to obtain cDNAs of genes that are expressed in cerebellar granule cells (GC), an antiserum was raised against GC isolated from mouse cerebella. Western blot analysis demonstrated that antibodies against multiple proteins were present and immunohistochemical analysis showed that at least some of these proteins were localized to cerebellar GC. The antiserum was used to screen an expression library derived from mouse cerebellar cDNA. Twenty-two granule cell antibody-positive (GCAP) clones were obtained. Of these, eight represented genes previously described and 14 were novel clones (not found in the GenBank database). In situ hybridization histochemistry showed that eight of the novel clones had moderate to strong expression in cerebellar GC and some of these clones were expressed also in the hippocampal formation. One such clone, GCAP-7, appears to represent a single-copy gene and the entire cDNA insert (2,688 bp) has been sequenced. The clone appears to consist primarily of the 3′ untranslated portion, including a poly(A) tail and polyadenylation signals,of a 5 kb transcript. The GCAP clones should be useful for future studies of molecular biology of GC in normal individuals and in inherited neurologic disease with GC degeneration.

Original languageEnglish (US)
Pages (from-to)183-191
Number of pages9
JournalMolecular Brain Research
Volume25
Issue number3-4
DOIs
StatePublished - Sep 1994

Keywords

  • Antiserum
  • cDNA expression library
  • Cerebellum
  • Granule cell
  • In situ hybridization
  • Novel clone
  • Western blot

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

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