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. ______________________________________________
Highlights
Paclitaxel (Taxol®), structurally, is one of the more complex members of the taxoid family of diterpenoid that are known as diverse group of secondary metabolites produced by yew (Taxus) species (Baloglu and Kingston, 1999)
Cloning of pTA-TXSHis in E. coli Top 10F was performed for reproduction of construct for further uses in transformation of Agrobacterium and Arabidopsis
For transformation of Agro-bacterium, a natural method entitled Triparental mating was used, in which the pillus of helper bacteria were commonly utilized to exchange the construct between E. coli and Agrobacterium (Bent, 2006)
Summary
Paclitaxel (Taxol®), structurally, is one of the more complex members of the taxoid family of diterpenoid that are known as diverse group of secondary metabolites produced by yew (Taxus) species (Baloglu and Kingston, 1999). It is the most dominant chemotherapeutic agent commonly used against various cancers such as breast, lung and ovarian cancers (Charles et al, 2001; Francis et al, 1995; Hata et al, 2004) as well as AIDS related Kaposi’s sarcoma (Gill et al, 1999). The grease-like mixture of three types of bacteria was spread on yeast extract broth (YEB) medium containing kanamycin sulfate (50 μg/ml) and rifampicin (25 μg/ml) This approach led to an appropriate primary selection of Agro-pTA-TXS-HIS.
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