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Promoter analysis of a medaka fish intestinal guanylyl cyclase gene
Author(s) -
Nakauchi Mina,
Suzuki Norio
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)00002-4
Subject(s) - gene , microbiology and biotechnology , fusion gene , biology , gene expression , luciferase , transcription (linguistics) , electrophoretic mobility shift assay , transfection , 5' flanking region , promoter , biochemistry , linguistics , philosophy
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.