| Summary: | 碩士 === 中山醫學大學 === 營養學研究所 === 94 === In this study, the search tool BLAST (Basic Local Alignment Search Tool), based on NCBI (National Center for Biotechnology Information) was used against nucleotide and protein databases. The analysis found a Dimeric Dihydrodiol Dihydrogenase (dimeric DD) of zebrafish (NCBI accession number BC081393/ AAH81393) and human (NCBI accession number NM_014475/NP_055290). The dimeric DD cDNA of zebrafish and human were successfully amplified with a continuous open reading frame of 1005 bps and 1005 bps encoding a protein of 334 and 334 amino acids with a calculated molecular mass of 36.740 kDa and 36.740 kDa respectively.
Zebrafish dimeric DD was cloned with a pGEM-T vector. To design a probe, which cdna of a antisense with DIG-UTP. Then, Zebrafish embryo were collected 24 hpf (hpf: hours post ertilization)、48 hpf、72 hpf and 96 hpf . Whole mount in situ hybridization analysis showed that dimeric DD is expressed in liver and pronephric ducts of 48 hpf、72 hpf and 96 hpf of Zebrafish embryo, not in 24 hpf of Zebrafish embryo. Therefore, this novel Dimeric Dihydrodiol Dihydrogenase is expressed in liver and pronephric ducts of larval Zebrafish not in more early stage.
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