J Integr Plant Biol. ›› 2002, Vol. 44 ›› Issue (12): 1450-1455.

• Research Articles • Previous Articles     Next Articles

Cloning, E. coli Expression and Molecular Analysis of a Novel Sesquiterpene Synthase Gene from Artemisia annua

LIU Yan, YE He-Chun and LI Guo-Feng   

Abstract:

A 1 886 bp full length sesquiterpene synthase (AaSES) cDNA was cloned from a high yield Artemisia annua L. strain 001 by a rapid amplification of cDNA end (RACE) strategy. AaSES is 59% identical to Artemisia cyclase cDNA clone cASC125, 50% identical to epicedrol synthase from A. annua, 48% identical to amorpha 4, 11 diene synthase from A. annua, 39% identical to the 5 epiaristolechene synthase from tabacco, 38% identical to vetispiradiene synthase from H. muticus, 41% identical to the δ cadinene synthase from cotton. The coding region of the cDNA was inserted into a procaryotic expression vector pET 30a and overexpressed in E. coli BL21(DE3). The yclase proteins extracted from bacterial culture were found largely in an insoluble protein fraction. AaSES expresses in leaves, stems and flowers, not in roots as indicated by Northern blotting analysis.

一个新高产青蒿倍半萜合酶基因的克隆、表达和分析
刘 彦 叶和春* 李国凤
(中国科学院植物研究所;中国科学院光合作用与环境分子生理重点实验室,北京1000093)

摘要: 用RACE方法从青蒿 (Artemisiaannua L .)高产株系 0 0 1中克隆了一个新的 1886bp的全长倍半萜合酶cDNA。克隆的倍半萜合酶氨基酸序列与烟草马兜铃烯合酶、莨菪岩兰螺旋二烯合酶、棉花杜松烯合酶的一致性分别为 39%、38%和 4 1% ;与青蒿柏木脑合酶、紫穗槐二烯合酶和一个推测的倍半萜合酶克隆cASC12 5的一致性为5 0 %、4 8%和 5 9%。cDNA编码区序列被克隆进原核表达载体pET-30a,并在大肠杆菌 (Escherichiacoli)BL2 1(DE3)中诱导表达 ,但过量表达的蛋白主要是以不溶性蛋白形式存在。Northernblotting分析表明此基因在茎、叶和花中表达 ,在根中没有表达。
关键词: 青蒿;倍半萜合酶;克隆

通讯作者。电话:+86(0)10 62591431-6249;E-mail: <hcye @ ns.ibcas.ac.cn>

Key words: Artemisia annua, sesquiterpene synthase, cloning

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