Journal of Molecular Neuroscience

, Volume 6, Issue 2, pp 109–119

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
Article

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.

Index Entries

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

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Copyright information

© Humana Press Inc 1995

Authors and Affiliations

  • Piet C. de Groen
    • 1
  • Bart J. L. Eggen
    • 2
  • Willem Hendrick Gispen
    • 3
  • Peter Schotman
    • 2
  • Loes H. Schrama
    • 2
  1. 1.Division of GastroenterologyMayo Clinic and FoundationRochester
  2. 2.Laboratory for Physiological ChemistryUtrecht UniversityUtrechtThe Netherlands
  3. 3.Department of Pharmacology, Rudolf Magnus Institute for NeurosciencesUtrecht UniversityUtrechtThe Netherlands

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