突变体
长春花
生物合成
吲哚试验
人口
化学
生物
生物化学
植物
基因
医学
环境卫生
作者
Mohammadamin Shahsavarani,Maisha Farzana,Vincenzo De Luca,Yang Qu
出处
期刊:Methods in molecular biology
日期:2022-01-01
卷期号:: 181-190
标识
DOI:10.1007/978-1-0716-2349-7_13
摘要
Decades of research on the medicinal plant Catharanthus roseus have led to the complete elucidation of the 29-step pathway for the biosynthesis of the anticancer drug vinblastine from geraniol and tryptophan precursors. Several approaches have been used to identify the enzymes involved in this iconic and remarkably complex pathway. This chapter describes the use of the classic ethyl methanesulfonate (EMS) mutagenesis to create a selfed M2 mutant population, which can be rapidly screened to select mutants with altered monoterpenoid indole alkaloid (MIA) biosynthesis with a simple, high-throughput thin-layer chromatography (TLC)-based screening strategy. This TLC-based MIA screening has led to the discovery and characterization of three enzymes responsible for vinblastine biosynthesis.
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