[Objective] This study aimed to clone Scrictosidine-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 RA...[Objective] This study aimed to clone Scrictosidine-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 CDS that encoded 536 amino acids with a calculated molecular mass of 61.0 kDa and an isoelectric point of 6.16. Bioinformatic analysis revealed that RvSGD shared high similarity with SGDs from Cantharanthus roseus and Rauvolfia serpentina at the amino acids; three conserved catalytic sites His-161, Glu-207 and Glu-419 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] The present study helps to understand more about the functions of the SGD gene at the level of molecular genetics, and provides new targets for molecular regulation of TIAs biosynthesis.展开更多
文摘[Objective] This study aimed to clone Scrictosidine-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 CDS that encoded 536 amino acids with a calculated molecular mass of 61.0 kDa and an isoelectric point of 6.16. Bioinformatic analysis revealed that RvSGD shared high similarity with SGDs from Cantharanthus roseus and Rauvolfia serpentina at the amino acids; three conserved catalytic sites His-161, Glu-207 and Glu-419 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] The present study helps to understand more about the functions of the SGD gene at the level of molecular genetics, and provides new targets for molecular regulation of TIAs biosynthesis.
