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目的:通过末端快速扩增(RACE)技术获得临安地区八角莲内闭联异松树脂醇二酯脱氢酶基因的全序列,探讨八角莲内异松树脂醇二酯脱氢酶基因的序列特点。方法:以浙江省临安地区八角莲为材料,根据同源序列法分别进行3′末端RACE和5′末端RACE,获得闭联异松树脂醇二酯脱氢酶基因的全序列。结果:以八角莲属的八角莲为材料获得闭联异松树脂醇二酯脱氢酶全基因序列为国际首次报道。此cDNA全长序列991bp,5′端非翻译区42bp,3′端非翻译区112bp。开放阅读框编码278个氨基酸,分子量为29253.3道尔顿,等电点为6.328。提交GeneBank序列号为EU573789。RT-PCR初步分析发现该酶的mRNA产物在茎内含量丰富。氨基酸序列比对表明闭联异松树脂醇二酯脱氢酶的物种差异性较大。结论:获得八角莲内闭联异松树脂醇二酯脱氢酶全基因。
OBJECTIVE: To obtain the full-length sequence of the homologue of isoprenoid dehydrogenase from the octagonal lotus in Lin’an through rapid end-amplification (RACE) technique. . METHODS: The full length sequence of the homologue of isorhamidin-2-phosphate dehydrogenase (DLDH) was obtained from Panax notoginseng Linn., Lin’an, Zhejiang Province using 3 ’terminal RACE and 5’ terminal RACE respectively. Results: It was the first time in the world to report the sequence of the homologue of cis-alditol resin diester dehydrogenase from the plant of the genus Octagus. The full-length cDNA of this cDNA is 991 bp in length, 42 bp in 5 ’untranslated region and 112 bp in 3’ untranslated region. The open reading frame encodes 278 amino acids with a molecular mass of 29253.3 Daltons and an isoelectric point of 6.328. Submit GeneBank serial number EU573789. Preliminary analysis by RT-PCR revealed that the mRNA product of this enzyme is abundant in the stem. Amino acid sequence alignment indicates that the species diversity of the different isoprenone diester dehydrogenases is greater. Conclusion: Obtained within the octagupidin homoserine alcohol diester dehydrogenase gene.