title: Post-transcriptional transfer of gamma-thio affinity label to RNA in isolated parsley nuclei.
abstract: As an alternative to soluble plant transcription systems, we examined the reinitiation capacity of isolated parsley nuclei. Nuclear chalcone synthase in vitro transcripts were affinity-labelled with gamma-thio-ATP, gamma-thio-GTP or beta-thio-ATP, and purified by chromatography on a mercury Sepharose affinity column. Primer extension and subsequent PCR amplification of these in vitro transcripts revealed gamma-thio-ATP-dependent, but no beta-thio-ATP-dependent, signals, although affinity labelling of overall in vitro transcripts still occurred with beta-thio-ATP. We conclude that the described plant nuclei reinitiated transcription non-specifically and that post-transcriptional transfer of the gamma-thio affinity label severely interfered with the detection of reinitiated transcripts.
authors: Max-Planck-Institut für Züchtungsforschung, Köln, FRG.
authors: P
authors: P
authors: Schweizer
authors: K
authors: K
authors: Hahlbrock
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D000345:Affinity Labels / Q000378:metabolism; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000378:metabolism; D005821:Genetic Techniques; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism*; D010944:Plants / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology*; D015533:Transcriptional Activation / Q000502:physiology
chemical_list: D000345:Affinity Labels; C022571:adenosine 5'-O-(3-thiotriphosphate); D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000255:Adenosine Triphosphate