We approach potyviral infection biology from different angles to gain insights into plant-virus interactions and their relevance for the infection.
On the Viikki campus we have access to specialized instruments and in-house expert services in for example proteomics, microscopy, next-generation sequencing and particle purification. Our plants are maintained in environmentally controlled greenhouses on campus.
Methods in a nutshell
- Pathosystems:
- potato virus A-Nicotiana benthamiana
- turnip mosaic virus-Arabidopsis thaliana
- Agroinfiltration-based gene transfer to host plants. We also initiate infections by agrobacterium-mediated transfer of infectious viral cDNA to the plants. Check out our agroinfiltration video to see how it is done.
- Quantitation of viral gene expression with a dual luciferase assay based on Renilla and firefly luciferase reporters
- Quantitative real time PCR and other methods to study mRNA levels
- Transient overexpression and RNA-silencing of host genes to study their role in potyviral infections
- Affinity purification and characterization of viral ribonucleoprotein complexes and host plant interaction partners in the infection context
- Recombinant protein expression and in vitro biochemical and structural analyses for studies on viral protein structures, functions and posttranslational modifications
- Polysome purification and ribosome profiling for studying host translation in the infection context.
- Microscopy: Epifluorescence and confocal microscopy to visualize in vivo localization and association of fluorescently tagged viral and host proteins. Electron microscopy for detailed imaging of infected cells and virus particles.
- Purification and enrichment of virus particles, infection-linked structures and organelles by ultracentrifugation.