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In-vitro transcription in tobacco chromatin.
- Source :
-
Planta [Planta] 1983 Oct; Vol. 159 (2), pp. 172-7. - Publication Year :
- 1983
-
Abstract
- A method is described to isolate transcriptionally active Nicotiana tabacum L. cv. Xanthi chromatin from suspension-cultured cells. Enzymatic preparation of protoplasts with solubilization of the plasma membrane, Triton X-100 and homogenization resulted in chromatin free from cellular debris. Incroporation of [(3)H]uridine triphosphate into RNA increased for more than 30 min at 30° C. Transcriptional activity was maximally stimulated at 10 mM MgCl2, 200 mM (NH4)2SO4 and 150 mM KCl. The in-vitro synthesized RNA was found to contain 3.8% polyadenylated RNA. The results of digestion studies with ribonuclease, heat and detergent inactivation studies, α-amanitin and actinomycin D inhibitor effects, as well as dependency on ribonucleotide triphosphates, showed that the activity recorded in the tobacco chromatin was bona-fide enzymatic RNA transcription. There also was marked stimulation of transcriptional activity when exogenous DNA was added to the assay mixture.
Details
- Language :
- English
- ISSN :
- 0032-0935
- Volume :
- 159
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Planta
- Publication Type :
- Academic Journal
- Accession number :
- 24258139
- Full Text :
- https://doi.org/10.1007/BF00392989