J Integr Plant Biol. ›› 2006, Vol. 48 ›› Issue (12): -.DOI: 10.1111/j.1744-7909.2006.00381.x

• Research Articles •    

Cloning, E. coli Expression and Molecular Analysis of Amorpha-4,11-Diene Synthase from a High-Yield Strain of Artemisia annua L.

Zhen-Qiu Li, Yan Liu, Ben-Ye Liu, Hong Wang, He-Chun Ye and Guo-Feng Li   

Abstract: Increasing demand of artemisinin in the treatment of malaria has placed substantial stress on the total artemisinin supplies world-wide, so more attention has been paid to increasing the content of artemisinin in the Artemisia annua L. plant. In this study, amorpha-4, 11-diene synthase (ADS) cDNA (ads1) and genomics gene (gads1) were cloned from a high-yield A. annua strain 001. The activity of ADS1 was confirmed by heterogeneous overexpression of ads1 and in vitro enzymatic incubation. Reverse transcript-polymerase chain reaction results demonstrated that ads1 expressed in leaves, flowers and young stems, but not in roots. This organ-specific expression pattern of ads1 is consistent with that of artemisinin accumulation in the plant. The gads1 has a complex organization including seven exons and six introns, and belongs to class III terpene synthase. DNA gel blotting revealed that the ADS gene has at least four copies in the genome of strain 001. The higher copy numbers might be one of the reasons for its high artemisinin content.(Author for correspondence. Tel: +86 (0)10 6283 6249; Fax: +86 (0)10 8259 1016; E-mail: hcye@ibcas.ac.cn)

Key words: amorpha-4,11-diene synthase, Artemisia annua, E. coli expression, molecular analysis.

Editorial Office, Journal of Integrative Plant Biology, Institute of Botany, CAS
No. 20 Nanxincun, Xiangshan, Beijing 100093, China
Tel: +86 10 6283 6133 Fax: +86 10 8259 2636 E-mail: jipb@ibcas.ac.cn
Copyright © 2022 by the Institute of Botany, the Chinese Academy of Sciences
Online ISSN: 1744-7909 Print ISSN: 1672-9072 CN: 11-5067/Q