[Objective] This study aimed to clone Scrietosidine-β-D-glucosidase (SGD) gene from Rauvolfia verticillata and analyze its characteristics. [Method] The full-length cDNA of SGD was cloned from R. verticillata with RACE technique. Then the expression levels in different tissues were analyzed with quantitative RT-PCR and the bioinformatic characteristics were also predicted. [Result] The full-length cDNA of RvSGD was 1 856 bp, containing a 1 608 bp ORF encoding 536 aniino acids with a calculated molecular mass of 61 kDa and an isoelectric point of 6. 16. Bioinformatic analysis revealed that RvSGD shared high similarity with SGDs from Vinca rosea and Rauvolfia serpenlina al the a-mino acids; three conserved catalytic sites His-161, Glu-207 and Glu419 were also presented in RvSGD. Quantitative RT-PCR showed that expression level of RvSGD was the highest in barks, followed by old leaves, roots, tender leaves and tender stems. [Conclusion J The present study will be helpful to understand more about the functions of the SGD gene at the level of molecular genetics, and provide new targets for molecular regulation of TIAs biosynthesis.%[目的]对萝芙木异胡豆苷-β-D-葡萄糖苷酶(SGD)基因进行克隆与分析.[方法]以萝芙木为材料,采用RACE技术,从萝芙木中克隆SGD的全长cDNA,并对其进行表达谱分析和相关生物信息学分析.[结果]萝芙木SGD基因cDNA全长1856 bp,包含1608 bp的开放阅读框,编码536个氨基酸,预测分子量为61 kDa,等电点p1为6.16;生物信息学分析显示,RvSGD与蛇根木SGD及长春花SGD的序列相似性分别高达90.1％和70.9％,RvSGD也具有SGD蛋白的3个保守氨基酸催化位点His-161、Glu-207和Glu-419;荧光定量PCR表明,RvSGD在树皮中表达量最高,其次是老叶、根、嫩叶和茎.[结论]RvSGD基因的克隆和功能分析有助于在分子水平上更好地了解这个基因在TIAs的生物合成中的作用,并为今后调控TIAs的生物合成提供了新的靶点.
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