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

M. Kambouris, L. C. Triarhou, Stephen Dlouhy, L. Sangameswaran, F. Luo, Bernardino Ghetti, M. E. Hodes

Research output: Contribution to journalArticle

6 Citations (Scopus)

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

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Gene Library
Complementary DNA
Clone Cells
Antibodies
Immune Sera
Genes
Polyadenylation
Nucleic Acid Databases
Nervous System Diseases
Cerebellum
In Situ Hybridization
Molecular Biology
Hippocampus
Proteins
Western Blotting
Databases
Messenger RNA

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

Cite this

Novel cDNA clones obtained by antibody screening of a mouse cerebellar cDNA expression library. / Kambouris, M.; Triarhou, L. C.; Dlouhy, Stephen; Sangameswaran, L.; Luo, F.; Ghetti, Bernardino; Hodes, M. E.

In: Molecular Brain Research, Vol. 25, No. 3-4, 1994, p. 183-191.

Research output: Contribution to journalArticle

Kambouris, M. ; Triarhou, L. C. ; Dlouhy, Stephen ; Sangameswaran, L. ; Luo, F. ; Ghetti, Bernardino ; Hodes, M. E. / Novel cDNA clones obtained by antibody screening of a mouse cerebellar cDNA expression library. In: Molecular Brain Research. 1994 ; Vol. 25, No. 3-4. pp. 183-191.
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