Cloning and promoter analysis of the human B-50/GAP-43 gene
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- de Groen, P.C., Eggen, B.J.L., Gispen, W.H. et al. J Mol Neurosci (1995) 6: 109. doi:10.1007/BF02736770
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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.