Abstract
Summary An improved method for the isolation of nuclei in a high yield from cultured tomato cells ( Lycopersicon peruvianum ) is described. The nuclei are active in RNA synthesis by run-on transcription. The transcripts were polydisperse in size without discrete peaks of RNA after polyacrylamide gel electrophoresis. Hybridization of the in vitro labelled RNA to DNA probes showed, that rRNA and mRNA genes were transcribed. Optimal conditions for a-amanitin sensitive (polymerase II) and insensitive (polymerase I) transcriptional activities were determined. Only the α-amanitin sensitive activity was stimulated by increasing concentrations of monovalent (ammonium sulfate) and divalent (magnesium) cations. Dot blot hybridization of the in vitro labelled transcripts to clones containing sequences for rRNA and two heat shock proteins has shown that the optimal specific transcription of rRNA but also of the mRNAs occured at lower salt concentrations. Under conditions of maximal RNA polymerase II activity (high salt concentrations) no increase in transcription of specific mRNA genes could be observed.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
