Plant Cell Reports

, Volume 19, Issue 5, pp 504–510

Preparation of transcriptionally active nuclei from etiolated Arabidopsis thaliana

Authors

  • K. M. Folta
    • Department of Biological Sciences Laboratory for Molecular Biology (m/c 567), 900 S. Ashland Avenue, Room 4150, University of Illinois at Chicago, Chicago, IL 60607, USA e-mail: lkaufman@uic.edu Fax: +312-4132691
  • L. S. Kaufman
    • Department of Biological Sciences Laboratory for Molecular Biology (m/c 567), 900 S. Ashland Avenue, Room 4150, University of Illinois at Chicago, Chicago, IL 60607, USA e-mail: lkaufman@uic.edu Fax: +312-4132691
Original Paper

DOI: 10.1007/s002990050764

Cite this article as:
Folta, K. & Kaufman, L. Plant Cell Reports (2000) 19: 504. doi:10.1007/s002990050764

Abstract

Despite its emergence as the plant model system, there are few reports that describe protocols for the isolation of functional nuclei from Arabidopsis thaliana and their use in nuclear run-on assays or in preparation of nuclear extracts. This is especially true for etiolated seedlings. Here we report conditions, optimized for use in Arabidopsis, which allow for the isolation of enriched fractions of functional nuclei from less than 2 g of etiolated or light-grown tissue. The nuclei are capable of incorporating 3H-UTP into TCA-insoluble RNA, and also incorporate 32P-CTP into transcripts that can subsequently be hybridized to specific filter-bound DNA target sequences. The functional nuclei are sensitive to the transcriptional inhibitors actinomycin D and α-amanitin, confirming that the transcription observed is both template dependent and relies on nuclear polymerase II.

ArabidopsisNucleiRun-onTranscriptionLight regulation

Copyright information

© Springer-Verlag Berlin Heidelberg 2000