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Molecular cloning and expression of retinoic-acid synthesizing enzyme raldh2 from Takifugu rubripes
Authors:Susumu Uji  Tohru Suzuki  Tadahide Kurokawa
Institution:aNational Research Institute of Aquaculture, Farming Biology Division, Fisheries Research Agency, 422-1, Nansei, Watarai, Mie 516-0193, Japan;bLaboratory of Bioindustrial Informatics, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai, Miyagi 981-8555, Japan
Abstract:Retinaldehyde dehydrogenases (raldhs) synthesize retinoic acid (RA), which is required for pattern formation and organogenesis during embryogenesis. To elucidate the common role of RA on vertebrate embryos, we first sought to clone a homologous gene to human raldh2 from fugu, Takifugu rubripes. We cloned a 1837 bp cDNA that encodes fugu raldh. The deduced amino acid sequence of the fugu raldh comprises 502 amino acids. The fugu Raldh showed highest sequence identity to zebrafish, Danio rerio, Raldh2 (79.9%). The fugu Raldh also showed high sequence identity to other vertebrate Raldh2: Xenopus laevis (77.2%), human (77.4%), mouse (74.3%) and chick (73.9%). Comparative genomic analysis showed that the gene arrangement around fugu raldh agreed with that of human raldh2. Fugu raldh mRNA was expressed through embryogenesis similarly to raldh2 in other vertebrates. These results and phylogenetic analyses suggest that pufferfish raldh is a fugu orthologue of other species' raldh2.
Keywords:cDNA  Embryogenesis  Genome  Raldh  Retinoic acid  RT-PCR  Takifugu rubripes  Vitamin A
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