Cloning and promoter analysis of the human B-50/GAP-43 gene

Piet C. de Groen, Bart J.L. Eggen, Willem Hendrick Gispen, Peter Schotman, Loes H. Schrama

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

We here report isolation of exon 1 and analysis of the human B-50 promoter. A human genomic λEMBL3 library was screened with a homologous PCR probe. Two independent clones were analyzed and partially sequenced: They contained up to 5 kb sequence upstream of the translation start site and approx 13 kb of intron 1 sequence. There was a high degree of homology between the rat and the human gene with 100% homology from -504 to -427, with respect to the translation start codon. However, relatively long GT and GA repeats as seen in the rat gene were absent. Various promoter-reporter constructs, containing 5.0 to 0.12 kb of the upstream region, were transfected into undifferentiated and neuroectodermally differentiated P19-EC. Two promoter activities were found. The minimal fragment with promoter activity still responsive to differentiation was the 0.22 kb construct, similar to rat promoter P2. We conclude that the human B-50 gene is expressed in a similar way to the rat B-50 gene, based on the presence of two transcripts, the high degree of homology between the rat and the human sequence, and the two promoter activities found in P19-EC cells.

Original languageEnglish (US)
Pages (from-to)109-119
Number of pages11
JournalJournal of Molecular Neuroscience
Volume6
Issue number2
DOIs
StatePublished - Jun 1 1995

Keywords

  • B-50/GAP-43 gene
  • Human
  • P19-EC cells
  • promoter

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

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    de Groen, P. C., Eggen, B. J. L., Gispen, W. H., Schotman, P., & Schrama, L. H. (1995). Cloning and promoter analysis of the human B-50/GAP-43 gene. Journal of Molecular Neuroscience, 6(2), 109-119. https://doi.org/10.1007/BF02736770