AI Article Synopsis
- Plant protoplasts are valuable tools for studying gene regulation and protein localization through quick screening methods.
- Protoplast transformation enables automated testing of plant promoters, particularly synthetic ones, like studies conducted with poplar mesophyll protoplasts.
- The study outlines a protocol for isolating poplar mesophyll protoplasts, transforming them, and analyzing images to identify promising synthetic promoters.
Article Abstract
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.
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10127045 | PMC |
| http://dx.doi.org/10.21769/BioProtoc.4660 | DOI Listing |
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