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Chapters
- 00:04Title
- 00:33Prepare Infiltration Suspension
- 02:11Perform Vacuum Infiltration
- 03:56Growth and Harvest of the Infiltrated Leaves and Pressurized Solvent Extraction
- 05:38Basic Ion Exchange Resin Treatment for Removal of Chlorophylls
- 06:58Results: Infiltration Coverage and Isolated Triterpene Product
- 07:49Conclusion
Transient heterologous expression of biosynthetic enzymes in Nicotiana benthamiana leaves can divert endogenous supplies of 2,3-oxidosqualene towards the production of new high-value triterpene products. Herein is described a detailed protocol for rapid (5 days) preparative-scale production of triterpenes and analogs utilizing this powerful plant-based platform.
Tags
Transient ExpressionNicotiana BenthamianaTriterpene ProductionPreparative ScaleAgrobacterium TumefaciensCo-infiltrationVacuum InfiltrationMMA BufferOD 600Preparative-scale AccessStructural CharacterizationMedicinally Relevant Biological ActivitySelective LB AgarSelective LB MediaCentrifugationResuspensionInfiltration Suspension
