Synthetic Promoter Screening Using Poplar Mesophyll Protoplast Transformation
- Univ. of Tennessee, Knoxville, TN (United States)
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States). Environmental Molecular Sciences Laboratory (EMSL)
- Univ. of California, Davis, CA (United States)
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.
- Research Organization:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER)
- Grant/Contract Number:
- AC05-76RL01830; SC0018347
- OSTI ID:
- 1988241
- Alternate ID(s):
- OSTI ID: 1988242
- Report Number(s):
- PNNL-SA-184602
- Journal Information:
- Bio-Protocol, Journal Name: Bio-Protocol Journal Issue: 8 Vol. 13; ISSN 2331-8325
- Publisher:
- Bio-protocol LLCCopyright Statement
- Country of Publication:
- United States
- Language:
- English
| Transcriptional landscape of highly lignified poplar stems at single-cell resolution | collection | January 2021 |