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Cloning and characterization of squalene synthase and cycloartenol synthase from Siraitia grosvenorii

机译:罗汉果角鲨烯角鲨烯合酶和环烯醇合酶的克隆与鉴定

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摘要

Mogrosides and steroid saponins are tetracyclic triterpenoids found in Siraitia grosvenorii. Squalene synthase (SQS) and cycloartenol synthase (CAS) are key enzymes in triterpenoid and steroid biosynthesis. In this study, full-length cDNAs of SgSQS and SgCAS were cloned by a rapid amplification of cDNA-ends with polymerase chain reaction (RACE-PCR) approach. The SgSQS cDNA has a 1254 bp open reading frame (ORF) encoding 417 amino acids, and the SgCAS cDNA contains a 2298 bp ORF encoding 765 amino acids. Bioinformatic analysis showed that the deduced SgSQS protein has two transmembrane regions in the C-terminal. Both SgSQS and SgCAS have significantly higher levels in fruits than in other tissues, suggesting that steroids and mogrosides are competitors for the same precursors in fruits. Combined in silico prediction and subcellular localization, experiments in tobacco indicated that SgSQS was probably in the cytoplasm or on the cytoskeleton, and SgCAS was likely located in the nucleus or cytosol. These results will provide a foundation for further study of SgSQS and SgCAS gene functions in S. grosvenorii, and may facilitate improvements in mogroside content in fruit by regulating gene expression.
机译:罗汉果苷和类固醇皂苷是罗汉果(Siraitia grosvenorii)中的四环三萜类化合物。角鲨烯合酶(SQS)和环戊烯醇合酶(CAS)是三萜和类固醇生物合成中的关键酶。在这项研究中,通过用聚合酶链反应(RACE-PCR)方法快速扩增cDNA末端,克隆了SgSQS和SgCAS的全长cDNA。 SgSQS cDNA具有1254 bp的开放阅读框(ORF),编码417个氨基酸,SgCAS cDNA包含2298 bp的ORF,编码765个氨基酸。生物信息学分析表明,推导的SgSQS蛋白在C末端具有两个跨膜区域。 SgSQS和SgCAS在水果中的含量均明显高于其他组织,这表明类固醇和罗汉果苷是水果中相同前体的竞争者。结合计算机预测和亚细胞定位,在烟草中的实验表明SgSQS可能在细胞质中或在细胞骨架上,而SgCAS可能位于细胞核或细胞质中。这些结果将为进一步研究罗汉果链球菌SgSQS和SgCAS基因功能提供基础,并可能通过调节基因表达来促进果实中罗汉果苷含量的提高。

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