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Characterization of an Ethanol-Inducible Promoter System in Catharanthus roseus Hairy Roots

机译:长春花毛状根中乙醇诱导型启动子系统的表征

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摘要

Efforts to engineer Catharanthus roseus hairy roots to produce commercially significant amounts of valuable compounds,such as the terpenoid indole alkaloids vinblastine and vincristine,require the development of tools to study the effects of overexpressing key metabolic and regulatory genes.The use of inducible promoters allows researchers to control the timing and level of expression of genes of interest.In addition,use of inducible promoters allows researchers to use a single transgenic line as both the control and experimental line,minimizing the problems associated with clonal variation.We have previously characterized the use of a glucocorticoid-inducible promoter system to study the effects of gene overexpression within the terpenoid indole alkaloid pathway on metabolite production.Here the feasibility of using an ethanol-inducible promoter within C.roseus hairy roots is reported.This ethanol-inducible promoter is highly sensitive to ethanol concentration with a concentration of 0.005% ethanol causing a 6-fold increase in CAT reporter activity after 24 h of induction.The ethanol-inducible CAT activity increased 24-fold over a 72-h induction period with 0.5% ethanol.
机译:努力改造长春花毛状根以产生商业上大量的有价值的化合物,例如萜类吲哚生物碱长春碱和长春新碱,需要开发工具来研究过表达关键代谢和调控基因的作用。诱导型启动子的使用使研究人员可以使用来控制目的基因表达的时间和水平。此外,诱导型启动子的使用使研究人员可以使用单个转基因系作为对照和实验系,从而最大限度地减少了与克隆变异相关的问题。糖皮质激素诱导型启动子系统研究萜类吲哚生物碱途径内基因过表达对代谢产物产生的影响。在此报道了在玫瑰果毛状毛根内使用乙醇诱导型启动子的可行性。对乙醇浓度敏感,浓度为诱导24小时后,0.005%的乙醇导致CAT报告子活性增加6倍。在0.5%乙醇诱导72小时的诱导期内,乙醇诱导的CAT活性增加了24倍。

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