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

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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.

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Authors and Affiliations

  1. Division of Gastroenterology, Mayo Clinic and Foundation, Rochester, MN

    Piet C. de Groen

  2. Laboratory for Physiological Chemistry, Utrecht University, Utrecht, The Netherlands

    Bart J. L. Eggen, Peter Schotman & Loes H. Schrama

  3. Department of Pharmacology, Rudolf Magnus Institute for Neurosciences, Utrecht University, Utrecht, The Netherlands

    Willem Hendrick Gispen

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  1. Piet C. de Groen
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  2. Bart J. L. Eggen
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  5. Loes H. Schrama
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de Groen, P.C., Eggen, B.J.L., Gispen, W.H. et al. Cloning and promoter analysis of the human B-50/GAP-43 gene. J Mol Neurosci 6, 109–119 (1995). https://doi.org/10.1007/BF02736770

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