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

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 using biotechnological 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.