Protocol for the purification of the plastid-encoded RNA polymerase from transplastomic tobacco plants

Protocol for the purification of the plastid-encoded RNA polymerase from transplastomic tobacco plants

Summary: 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...

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Bibliographic Details
Main Authors: Xiao-Xian Wu, Fan Li, Chuxia Zhu, Shu-Yi Sun, Wen-Hui Mu, Stephanie Ruf, Ralph Bock, Yu Zhang, Fei Zhou
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:STAR Protocols
Subjects:
Molecular Biology
Plant sciences
Protein Biochemistry
Structural Biology
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724006932
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Summary:Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667

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