Wang Gui-Ling, Huang Dong-Yang
Key Laboratory of Ministry of Public Health, Department of Cell Biology, China Medical University, Shenyang 110001, China.
Yi Chuan Xue Bao. 2004 Apr;31(4):403-10.
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.
从牛肝脏中提取总RNA。根据人、小鼠和兔烟酰胺腺嘌呤二核苷酸磷酸（NADP(H)）依赖性视黄醇脱氢酶/还原酶（NRDR）基因的cDNA序列，设计并合成了基因特异性引物。在本研究中，我们通过cDNA末端快速扩增（RACE）和逆转录-聚合酶链反应（RT-PCR）方法克隆了牛肝脏NRDR的全长cDNA。牛NRDR cDNA为1266 bp，其开放阅读框（ORF）与其他NRDR cDNA一样，为783 bp，编码260个氨基酸残基。牛NRDR在氨基酸序列上与人、小鼠和兔的NRDR具有同一性。它在C末端含有短链脱氢酶/还原酶（SDR）保守基序和过氧化物酶体靶向信号（PTS1）序列。总之，利用RACE方法成功克隆了牛NRDR cDNA，并提交至GenBank（AF487454），利用生物信息学对其核苷酸序列和推导的氨基酸序列进行了分析，该基因属于一个新的过氧化物酶体SDR超家族，在视黄酸合成的限速步骤中起重要作用。这是首次报道表明哺乳动物过氧化物酶体的SDR参与类视黄醇代谢，为进一步研究该蛋白的生物学功能和视黄酸生物合成提供了可靠的基础。
