Cargando…

Synthetic Promoter Screening Using Poplar Mesophyll Protoplast Transformation

Plant protoplasts are useful to study both transcriptional regulation and protein subcellular localization in rapid screens. Protoplast transformation can be used in automated platforms for design-build-test cycles of plant promoters, including synthetic promoters. A notable application of protoplas...

Descripción completa

Detalles Bibliográficos
Autores principales: Yang, Yongil, Shao, Yuanhua, Chaffin, Timothy A., Ahkami, Amir H., Blumwald, Eduardo, Stewart, C. Neal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bio-Protocol 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10127045/
https://www.ncbi.nlm.nih.gov/pubmed/37113331
http://dx.doi.org/10.21769/BioProtoc.4660
Descripción
Sumario:Plant protoplasts are useful to study both transcriptional regulation and protein subcellular localization in rapid screens. Protoplast transformation can be used in automated platforms for design-build-test cycles of plant promoters, including synthetic promoters. A notable application of protoplasts comes from recent successes in dissecting synthetic promoter activity with poplar mesophyll protoplasts. For this purpose, we constructed plasmids with TurboGFP driven by a synthetic promoter together with TurboRFP constitutively controlled by a 35S promoter, to monitor transformation efficiency, allowing versatile screening of high numbers of cells by monitoring green fluorescent protein expression in transformed protoplasts. Herein, we introduce a protocol for poplar mesophyll protoplast isolation followed by protoplast transformation and image analysis for the selection of valuable synthetic promoters. Graphical overview [Image: see text]