Molecular cloning and expression of CCAAT/enhancer binding proteins in Japanese flounder Paralichthys olivaceus.
The findings of this study represent the first report, to the authors' knowledge, of CCAAT/enhancer binding protein (C/EBP) cDNA sequence in a fish species. C/EBP epsilon of Japanese flounder was 1861 bp in length (ORF of 822 bp) encoding for 274 amino acids, with a calculated molecular weight of 30 kDa. Japanese flounder C/EBP beta was found to be 1561 bp in length (ORF of 1041 bp), encoding for 347 amino acids and a calculated molecular weight of 39 kDa. These genes were expressed in various fish organs, tissues and secretions. C/EBP epsilon was detected by Northern blot from total RNA of head and posterior kidney, heart and spleen. However, RT-PCR also detected C/EBP epsilon in brain, spleen and peritoneal cavity fluid and peripheral blood leucocyte cDNA. C/EBP beta was detected by Northern blot analysis in the head and posterior kidney, spleen, intestine, liver, brain, heart, gill and testis and further found by RT-PCR to be detected in mucus, peritoneal cavity fluid, peripheral blood leucocytes and eye cDNA. Phylogenetic analysis placed the Japanese flounder C/EBP beta within the same cluster as previously reported C/EBP beta sequences. However, Japanese flounder C/EBP epsilon sequence data were not found to cluster with the three reported mammalian C/EBP epsilon sequences currently available. Understanding C/EBP transcriptional gene control in commercially important fish species may lead to a better control of disease.
Tucker
Carl S
CS
Laboratory of Genetics and Biochemistry, Tokyo University of Fisheries, Konan 4-5-7, Minato-ku, Tokyo 108-8477, Japan.
Hirono
Ikuo
I
Aoki
Takashi
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D022762:CCAAT-Enhancer-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D005432:Flounder / Q000145:classification / Q000276:immunology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction
D022762:CCAAT-Enhancer-Binding Proteins; D018076:DNA, Complementary