[Cloning and analysis of NADP(H) -dependent retinol dehydrogenase/reductase gene: a novel member of short-chain dehydrogenase/reductase].
The total RNA was purified from bovine liver. According to the cDNA sequences of human, mouse and rabbit NADP(H)-dependent retinol dehydrogenase/reductase (NRDR) gene, the gene-specific primers were designed and synthesized. In this study, we cloned the full-length cDNA of bovine liver NRDR by rapid amplification of cDNA ends (RACE) and RT-PCR methods. The bovine NRDR cDNA is 1 266 bp and the ORF, like other NRDR cDNAs, is 783 bp, encoding 260 amino acid residues. The bovine NRDR exhibited the identity in amino acid sequence to those of human, mouse and rabbit NRDR. It contains short-chain dehydrogenase/reductase (SDR) conservative motif and the peroxisomal targeting singal (PTS1) sequence at their C-terminal. In conclusion, the bovine NRDR cDNA was successfully cloned with RACE methods and submitted to GenBank(AF487454), whose nucleotide sequence and the deduced amino acid sequence were analyzed with Bioinformatics, that belongs to a novel peroxisomal SDR superfamily and plays an important role in the rate-limiting step of synthesizing retinoic acid. This is the first report suggesting the SDR participation of mammalian peroxisomers in retinoid metabolism and it provides a reliable foundation to further investigate the biological function of this protein and retinoic acid biosynthesis.
Key Laboratory of Ministry of Public Health, Department of Cell Biology, China Medical University, Shenyang 110001, China. wanggl2000@163.com
Gui-Ling
GL
Wang
Dong-Yang
DY
Huang
D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D015723:Gene Library; D008969:Molecular Sequence Data; D009249:NADP / Q000378:metabolism*
D018076:DNA, Complementary; D009249:NADP; D000429:Alcohol Oxidoreductases; C022520:retinol dehydrogenase