Promoter analysis of a medaka fish intestinal guanylyl cyclase gene.
We characterized the promoter activity of a medaka fish intestinal guanylyl cyclase gene, OlGC6, by assay of enzyme activity in response to various promoter-luciferase fusion gene constructs introduced into CACO-2 cells and medaka fish embryos. A transient transfection assay of the various fusion gene constructs showed that the nucleotides between -98 and -89 in the 5'-flanking region of the OlGC6 gene are essential for transcription of the OlGC6 gene in CACO-2, and that the OlGC6 gene fragment between -98 and +50 is sufficient to drive gene expression in the medaka fish intestine. An electrophoretic mobility shift assay and ultraviolet (UV) cross-linking experiments demonstrated that a nuclear protein from CACO-2 cells and the adult medaka fish intestinal cells binds specifically to the AGACCTTTGC nucleotides in the regulatory element.
Nakauchi
Mina
M
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
Suzuki
Norio
N
D024506:5' Flanking Region; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D019556:COS Cells; D018938:Caco-2 Cells; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D006162:Guanylate Cyclase / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000201:enzymology*; D009990:Oryzias / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D020218:Response Elements; D015533:Transcriptional Activation
D004268:DNA-Binding Proteins; D006162:Guanylate Cyclase