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Charakterisierung der Promotoraktivität des ovinen PrP Gens in N2a Neuroblastom‐ und ovinen fetalen Gehirnzell‐Linien
Author(s) -
O'Neill G. T.,
Donnelly K.,
Marshall E.,
Cairns D.,
Goldmann W.,
Hunter N.
Publication year - 2003
Publication title -
journal of animal breeding and genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 51
eISSN - 1439-0388
pISSN - 0931-2668
DOI - 10.1046/j.1439-0388.2003.00381.x
Subject(s) - gene , exon , biology , microbiology and biotechnology , reporter gene , promoter , coding region , regulatory sequence , transcription (linguistics) , transfection , gene expression , genetics , linguistics , philosophy
Summary In this study, we have analysed the DNA sequence and promoter activity of a 522 base pair non‐coding region of the sheep PrP gene representative of exon I and 440 bases upstream of the exon I start site. DNA sequence analyses of the PrP promoter region from Cheviot sheep identified a C/G dimorphism that created a SP1 transcription factor binding site. Additionally, a new T/C dimorphism was identified within Motif 1, one of four highly conserved sequence motifs common to all PrP promoter gene sequences. In vitro gene promoter activity was analysed by chloramphenicol acetyl transferase (CAT) reporter gene transfection assays in murine N2a cells and sA80BR, a permanently transformed cell line derived from a foetal Cheviot sheep brain. The minimal PrP gene sequences required for CAT reporter gene expression in N2a and sheep sA80BR cells were found to be 75 bases and 131 bases proximal to the exon I start site, respectively. No significant differences in PrP gene promoter activity was observed following the sequential deletion of the four conserved motifs though an initial loss of promoter activity was observed following the deletion of the first 99 bases of the promoter sequence.