We examined the spatial expression pattern of medaka fish (Oryzias latipes) soluble guanylate cyclase α1 and β1 subunit genes, OlGCS-α1 and OIGCS-β1, and characterized the 5′-flanking region required for expression of both genes by introducing various promoter-luciferase fusion-gene constructs into COS-1 cells and medaka fish embryos. The OlGCS-α1 and OIGCS-β1 gene transcripts were detected in whole brain and kidney in 7-day and 9-day embryos. Primer-extension analysis demonstrated that there were no differences among various adult organs (brain, eye, kidney, ovary and testis) in the transcription start site of the OIGCS-α1 and OIGCS-β1 genes. Neither gene contained the functional TATA box within its 5′-flanking region, and the basal promoter activity was found between nucleotides +33 and +42 in the OIGCS-α1 gene and between nucleotides + 146 and + 155 in the OIGCS-β1 gene. In the assay of medaka fish embryos, the 5′-flanking region of the OIGCS-β1 gene exhibited lower promoter activity than that of the OIGCS-α1 gene. In the experiments on dual-luciferase fusion-gene constructs, the 5′-flanking region of the OIGCS-α1 gene connected to the 5′-flanking region of the OIGCS-β1 gene was introduced into medaka fish embryos, and the 5′-flanking regions of both subunit genes were shown to mutually influence each other's promoter activity.
- Co-ordinated transcription
- Dual-luciferase assay
- Reporter fusion-gene construct
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