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Examining the transcriptional response of overexpressing anthranilate synthase in the hairy roots of an important medicinal plant Catharanthus roseus by RNA-seq

机译:通过RNA序列检查重要药用植物长春花的毛状根中过表达邻氨基苯甲酸合酶的转录反应

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摘要

BackgroundClinically important anti-cancer drugs vinblastine and vincristine are solely synthesized by the terpenoid indole alkaloid (TIA) pathway in Catharanthus roseus. Anthranilate synthase (AS) is a rate-limiting enzyme in the TIA pathway. The transgenic C. roseus hairy root line overexpressing a feedback insensitive ASα subunit under the control of an inducible promoter and the ASβ subunit constitutively was previously created for the overproduction of TIAs. However, both increases and decreases in TIAs were detected after overexpressing ASα. Although genetic modification is targeted to one gene in the TIA pathway, it could trigger global transcriptional changes that can directly or indirectly affect TIA biosynthesis. In this study, Illumina sequencing and RT-qPCR were used to detect the transcriptional responses to overexpressing AS, which can increase understanding of the complex regulation of the TIA pathway and further inspire rational metabolic engineering for enhanced TIA production in C. roseus hairy roots.
机译:背景临床上重要的抗癌药物长春碱和长春新碱是通过长春花中的萜类吲哚生物碱(TIA)途径合成的。邻氨基苯甲酸合酶(AS)是TIA途径中的限速酶。先前创建用于诱导TIA过量生产的在诱导型启动子的控制下过表达反馈不敏感的ASα亚基和组成型ASβ亚基的转基因玫瑰花毛状根系。但是,过表达ASα后,TIA的增加和减少均被检测到。尽管基因修饰针对的是TIA途径中的一个基因,但它可能触发可直接或间接影响TIA生物合成的全局转录变化。在这项研究中,Illumina测序和RT-qPCR被用于检测对过表达AS的转录反应,这可以增加对TIA途径复杂调控的理解,并进一步激发合理的代谢工程,以提高玫瑰色梭菌毛状根中TIA的产生。

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