Protocol for the purification of the plastid-encoded RNA polymerase from transplastomic tobacco plants
The plastid-encoded RNA polymerase (PEP) plays an essential role in the transcription of the chloroplast genome. Here, we present a strategy to purify the transcriptionally active protein complex from transplastomic tobacco (Nicotiana tabacum) lines in which one of the PEP core subunits is fused to an epitope tag. We describe experimental procedures for designing transformation constructs for PEP purification, selection, and analysis of transplastomic tobacco plants. We then detail the steps for purifying PEP from the transplastomic tobacco leaves. For complete details on the use and execution of this protocol, please refer to Wu et al.<sup>1</sup>.
Authors
- Chuxia Zhu (ORCID: https://orcid.org/0000-0002-5980-9854)
- Xiaoxian Wu (ORCID: https://orcid.org/0000-0002-1903-8151)
- Wenhui Mu
- Ralph Bock (ORCID: https://orcid.org/0000-0001-7502-6940)
- Stephanie Ruf (ORCID: https://orcid.org/0000-0002-9469-5944)
- Yu Zhang (ORCID: https://orcid.org/0000-0002-1778-8389)
- Fan Li (ORCID: https://orcid.org/0009-0009-6274-6100)
- Fei Zhou (ORCID: https://orcid.org/0000-0002-6197-3629)
- Sun Shu-yi
Institutions
- Huazhong Agricultural University (CN)
- Max Planck Institute of Molecular Plant Physiology (DE)
- Center for Excellence in Molecular Plant Sciences (CN)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-01-10
- DOI
- https://doi.org/10.1016/j.xpro.2024.103528
- Citations
- 1
- Primary Topic
- Photosynthetic Processes and Mechanisms
- Type
- article
- Field-Weighted Citation Impact
- 0.81
Funders
- National Key Research and Development Program of China