An Engineered Heat-Inducible Expression System for the Production of Casbene in Nicotiana benthamiana

Plants respond to heat stress by producing heat-shock proteins. These are regulated promoters containing regulatory elements, which can be harnessed control protein expression both temporally and spatially. In this study, we designed heat-inducible produce the diterpene casbene in Nicotiana benthamiana, through a multi-step metabolic pathway. To potentially increase gene transcription, coupled elements from Arabidopsis thaliana Hsp101 or Glycine max GmHsp17.3-B promoters, CAAT TATA boxes CaMV 35S, 5′UTR tobacco mosaic virus. The resulting four chimeric fused green fluorescent (GFP) reporter showed that variant Ara2 had strongest signal after shock. We next created 4-gene cassette driven promoter allow for exogenous synthesis of transformed multigene construct along with selectable marker into benthamiana. Metabolic analysis on transgenic lines revealed continuous outperforms shock, up 1 μg/mg DW detected 32 h uninterrupted 40 °C heat. results demonstrate potential as synthetic biology tools metabolite production plants.