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OALib Journal期刊
ISSN: 2333-9721
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曼地亚红豆杉ipi基因克隆与功能鉴定
, PP. 621-626
Keywords: 曼地亚红豆杉,紫杉醇,异戊烯基焦磷酸异构酶,功能验证
Abstract:
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(isopentenyldiphosphateisomerase,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(genbank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cdna全长1232bp,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:tmipi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。
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