Cloning and Expression of cDNA Encoding Key Enzymes (DXR,SLS,G10H and STR) in Terpene Indole Alkaloids Biosynthesis Pathway from Catharanthus roseus

doi:10.7525/j.issn.1673-5102.2007.05.013

Bulletin of Botanical Research ›› 2007, Vol. 27 ›› Issue (5): 564-568.doi: 10.7525/j.issn.1673-5102.2007.05.013

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Cloning and Expression of cDNA Encoding Key Enzymes (DXR,SLS,G10H and STR) in Terpene Indole Alkaloids Biosynthesis Pathway from Catharanthus roseus

HAN Mei;ZHAO Bo;AN Zhi-Gang;Thomas Rausch;ZU Yuan-Gang*

1.Key Laboratory of Forest Plant Ecology,Ministry of Education,Northeast Forestry University,Harbin　150040 2.Institute of Plant Science,Heidelberg University,Heidelberg,Germany

Received:1900-01-01 Revised:1900-01-01 Online:2007-09-20 Published:2007-09-20
Contact: ZU Yuan-Gang
Supported by:

Abstract

Abstract: cDNAs encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR)，Secologanin synthase (SLS)，Geraniol 10-hydroxylase(G10H) and Strictosidine synthase (STR) were obtained from the leaves of Catharanthus roseus with 2-step gateway PCR. These cDNA clones were inserted into the GATEWAY donor vector pDONR201 at the corresponding sites by BP clonase and followed by sequencing analysis,respectively. Thereafter,the entry clones were introduced into the GATEWAY destination vector pETG10A fused to 6xHIS-tag by LR clonase. Overexpression of recombinant fusion proteins DXR,SLS and STR were initiated after IPTG induction. These recombinant fusion proteins are now available for developing the antisera.

Key words: Catharanthus roseus, terpene indole alkaloids, biothsynthesis pathway, gateway clone

CLC Number:

HAN Mei;ZHAO Bo;AN Zhi-Gang;Thomas Rausch;ZU Yuan-Gang*. Cloning and Expression of cDNA Encoding Key Enzymes (DXR,SLS,G10H and STR) in Terpene Indole Alkaloids Biosynthesis Pathway from Catharanthus roseus[J]. Bulletin of Botanical Research, 2007, 27(5): 564-568.

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Cloning and Expression of cDNA Encoding Key Enzymes (DXR,SLS,G10H and STR) in Terpene Indole Alkaloids Biosynthesis Pathway from Catharanthus roseus

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