African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Full Length Research Paper

Cloning of taxadiene synthase gene into Arabidopsis thaliana (ecotype Columbia-0)

Sajjad Khani1,5, Mohammad Mehdi Sohani2, Nasser Mahna3, Jaleh Barar1, Mohammad Saeed Hejazi4, Hossein Nazemieh1,4, Sina Atashpaz1,5, Mohammad Reza Dadpour1,3 and Yadollah Omidi1,5*
1Research Centre for Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. 2Department of Agricultural Biotechnology, Faculty of Agriculture, Guilan University, Rasht, Iran. 3Department of Horticultural Sciences, Faculty of Agriculture, University of Tabriz, Tabriz, Iran. 4Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. 5School of Advanced Biomedical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Email: [email protected]

  •  Accepted: 24 November 2009
  •  Published: 22 March 2010

Abstract

Paclitaxel (Taxol), a complex diterpenoid, produced by yew tree (Taxus sp.) is the most important chemotherapeutic agent that is widely used against a variety of malignancies such as ovarian and breast cancers. However, destructive methods for its production from natural resources together with currently used low-yielding industrial production systems via total synthesis or semi-synthesis have led researchers to invent a robust alternative biological production system usingbiotechnological approaches. The first committed step in taxol biosynthesis pathway is the production of taxadiene from geranylgeranyl diphosphate (GGPP) catalyzed by the plastid-localized enzyme taxadiene synthase (TXS). In this research, an attempt was made to evaluate the effects of the first critical enzyme in the taxol biosynthesis pathway on Arabidopsis plant through the expression of taxadiene synthase gene under the control of a dexamethasone-inducible promoter.To achieve this goal, Arabidopsis plants (ecotype Columbia-0) were transformed with the construct pTA-TXS-His via floral dip method using Agrobacterium tumefaciens AGL1. The transformed plants were confirmed using the PCR reaction amplifying an 800 bp fragment of the cloned gene. Upon these findings, a proposal was made that biotechnological strategies could be utilized for the production of taxol components. 

 

Key words: Paclitaxel, arabidopsis, taxadiene synthase, geranylgeranyl diphosphate.

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