Cloning and promoter analysis of the human B-50/GAP-43 gene
Rent the article at a discountRent now
* Final gross prices may vary according to local VAT.Get Access
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.
- Cloning and promoter analysis of the human B-50/GAP-43 gene
Journal of Molecular Neuroscience
Volume 6, Issue 2 , pp 109-119
- Cover Date
- Print ISSN
- Online ISSN
- Humana Press
- Additional Links
- B-50/GAP-43 gene
- P19-EC cells
- Industry Sectors
- Author Affiliations
- 1. Division of Gastroenterology, Mayo Clinic and Foundation, Rochester, MN
- 2. Laboratory for Physiological Chemistry, Utrecht University, Utrecht, The Netherlands
- 3. Department of Pharmacology, Rudolf Magnus Institute for Neurosciences, Utrecht University, Utrecht, The Netherlands