青蒿
甲戊酸途径
生物
青蒿素
毛状体
生物化学
法尼基二磷酸合酶
生物合成
杰马克林
异源表达
倍半萜
互补DNA
基因
萜类
ATP合酶
植物
恶性疟原虫
重组DNA
免疫学
疟疾
作者
Cinzia M. Bertea,Alessandra Voster,Francel Verstappen,Massimo E. Maffei,Jules Beekwilder,Harro J. Bouwmeester
标识
DOI:10.1016/j.abb.2006.02.026
摘要
Artemisia annua (Asteraceae) is the source of the anti-malarial compound artemisinin. To elucidate the biosynthetic pathway and to isolate and characterize genes involved in the biosynthesis of terpenoids including artemisinin in A. annua, glandular trichomes were used as an enriched source for biochemical and molecular biological studies. The sequencing of 900 randomly selected clones from a glandular trichome plasmid cDNA library revealed the presence of many ESTs involved in isoprenoid biosynthesis such as enzymes from the methylerythritol phosphate pathway and the mevalonate pathway, amorpha-4,11-diene synthase and other sesquiterpene synthases, monoterpene synthases and two cDNAs showing high similarity to germacrene A synthases. Full-length sequencing of the latter two ESTs resulted in a 1686-bp ORF encoding a protein of 562 aa. Upon expression in Escherichia coli, the recombinant protein was inactive with geranyl diphosphate, but catalyzed the cyclization of farnesyl diphosphate to germacrene A. These results demonstrate the potential of the use of A. annua glandular trichomes as a starting material for studying isoprenoid biosynthesis in this plant species.
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