Characterization of promoter activities of four different Japanese flounder promoters in transgenic zebrafish.
An important consideration in transgenic research is the choice of promoter for regulating the expression of a foreign gene. In this study several tissue-specific and inducible promoters derived from Japanese flounder Paralichthys olivaceus were identified, and their promoter activity was examined in transgenic zebrafish. The 5' flanking regions of the Japanese flounder complement component C3, gelatinase B, keratin, and tumor necrosis factor (TNF) genes were linked to green fluorescence protein (GFP) as a reporter gene. The promoter regulatory constructs were introduced into fertilized zebrafish eggs. As a result we obtained several stable transgenic zebrafish that displayed green fluorescence in different tissues. Complement component C3 promoter regulated GFP expression in liver, and gelatinase B promoter regulated it in the pectoral fin and gills. Keratin promoter regulated GFP expression in skin and liver. TNF gene promoter regulated GFP expression in the pharynx and heart. TNF promoter had lipoplysaccharide-inducible activity, such that when transgenic embryos were immersed lipopolysaccharide, GFP expression increased in the epithelial tissues. These 4 promoters regulated the expression of GFP in different patterns in transgenic zebrafish.
Yazawa
Ryosuke
R
Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato, Tokyo 108-8477, Japan.
Hirono
Ikuo
I
Aoki
Takashi
T
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D001483:Base Sequence; D003176:Complement C3 / Q000235:genetics; D017931:DNA Primers; D005432:Flounder / Q000235:genetics*; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007633:Keratins / Q000235:genetics; D008099:Liver / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012867:Skin / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics; D015027:Zebrafish
D003176:Complement C3; D017931:DNA Primers; D014409:Tumor Necrosis Factor-alpha; D049452:Green Fluorescent Proteins; D007633:Keratins; D020780:Matrix Metalloproteinase 9