Abstract
In order to study the effects oftrans-acting factors oncis-acting elements in plant genes anin vitro reinitiating nuclear transcription system is needed. Here we report that run-on experiments with nuclei isolated from 2,4-D-treated auxin-starved early-stationary-phase cells of tobacco clearly show reinitiation of transcription of specific 2,4-D-induced genes. Using γ-thio nucleotides and [α-32P]-UTP we were able to demonstrate the presence of reinitiated labelled specific RNAs after isolation on a mercury-agarose affinity column. Addition of heparin as an inhibitor blocked this reinitiation. In a primer extension assay we found that the new transcripts initiate at approximately the same site as usedin vivo.
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Abbreviations
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- Tris:
-
2-amino-2-(hydroxymethyl)-1,3-propanediol
- BSA:
-
bovine serum albumin
- SDS:
-
sodium dodecyl sulphate ME, mercaptoethanol
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Mennes, A.M., Quint, A., Gribnau, J.H. et al. Specific transcription and reinitiation of 2,4-D-induced genes in tobacco nuclei. Plant Mol Biol 18, 109–117 (1992). https://doi.org/10.1007/BF00018461
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DOI: https://doi.org/10.1007/BF00018461