Yamada K, Oura Y, Mori H, Yamaki S
Graduate School of Bioagricultural Sciences, Nagoya University, Japan.
Plant Cell Physiol. 1998 Dec;39(12):1375-9. doi: 10.1093/oxfordjournals.pcp.a029345.
Partial amino acid sequences of NAD-dependent sorbitol dehydrogenase (NAD-SDH) were used to identify a full-length cDNA from apple fruit. This clone consisted of 1,433 bp containing an open reading frame of 1,137 bp that could code for a polypeptide with 379 amino acids. To our knowledge, this is the first report about cloning of NAD-SDH cDNA from a plant source. The deduced amino acids from cDNA revealed 43.7% identity to human NAD-SDH. The activity of this enzyme to convert sorbitol to fructose with the reduction of NAD was certified by the fusion protein of this clone expressed in Escherichia coli. Northern blot analysis showed that the mRNA was expressed in matured apple fruit.
利用NAD依赖的山梨醇脱氢酶（NAD-SDH）的部分氨基酸序列从苹果果实中鉴定出一个全长cDNA。该克隆由1433个碱基对组成，包含一个1137个碱基对的开放阅读框，可编码一个含有379个氨基酸的多肽。据我们所知，这是关于从植物来源克隆NAD-SDH cDNA的首次报道。从cDNA推导的氨基酸与人类NAD-SDH有43.7%的同一性。该克隆在大肠杆菌中表达的融合蛋白证实了这种酶将山梨醇转化为果糖并伴随NAD还原的活性。Northern印迹分析表明，该mRNA在成熟苹果果实中表达。
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