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紫杉醇(taxol)是从裸子植物红豆杉中提取的具有抗癌作用的天然药物。异戊烯基焦磷酸异构酶(isopentenyl diphosphate isomerase,IPI)是紫杉醇生物合成途径上催化IPP和二甲基烯丙基焦磷酸(DMAPP)之间进行可逆转化的异构酶。本研究从曼地亚红豆杉中首次克隆了编码IPI蛋白的基因序列(Gen Bank登录号为KP970677),并对其进行详细的生物信息学分析。研究表明,本研究所获得的IPI基因c DNA全长1 232 bp,编码233个氨基酸,包含Nudix保守结构域,与其他物种的IPI具有高度同源性;系统进化分析表明,植物IPI被分为被子植物和裸子植物两大类群,本文所研究的红豆杉IPI属于裸子植物IPI类群,该结果与红豆杉属于裸子植物这一事实相符;Southern杂交结果表明:Tm IPI是紫杉醇等萜类生物合成途径IPI家族成员。功能验证结果表明:在大肠杆菌中过量表达IPI推动代谢流向下游流动,促进β-胡萝卜素的生物合成,说明IPI在紫杉醇等萜类相关产物生物合成中是一个重要的调节因子。
Taxol is a natural drug with anti-cancer activity extracted from the gymnospermous yew. Isopentenyl diphosphate isomerase (IPI) is an isomerase that catalyzes the reversible transformation between IPP and dimethylallyl pyrophosphate (DMAPP) on the paclitaxel biosynthetic pathway. In this study, the gene sequence encoding the IPI protein was first cloned from Taxus media, with Gen Bank accession number KP970677, and detailed bioinformatics analysis was performed. Studies have shown that the cDNA of IPI gene obtained in this study is 1 232 bp in length and encodes 233 amino acids. It contains a Nudix conserved domain and shares a high degree of homology with IPI of other species. Phylogenetic analysis shows that plant IPI is divided into The angiosperms and gymnosperms are two major groups. The IPI of Taxus chinensis studied in this paper belongs to the IPI group of gymnosperms. This result is consistent with the fact that Taxus is a gymnosperm plant; Southern blot results indicate that Tm IPI is an anthraquinone biosynthesis pathway such as paclitaxel. Members of the IPI family. The results of functional verification showed that overexpression of IPI in E. coli promoted the downstream flow of metabolic pathways and promoted the biosynthesis of β-carotene, indicating that IPI is an important regulatory factor in the biosynthesis of terpenoid related products such as paclitaxel.