title: Preferential transcription of the ovalbumin gene in isolated hen oviduct nuclei by RNA polymerase B.
abstract: The synthesis of ovalbumin mRNA sequences was studied in isolated nuclei from hen oviduct. Two different methods of analysis were used to distinguish in vitro synthesized from preexisting mRNA sequences: (i) Mercurated ribonucleotides were used for in vitro RNA synthesis, and the newly synthesized RNA was purified by chromatography on sulfhydryl-agarose and hybridized to radioactive ovalbumin cDNA. (ii) [3H]UTP was used to label the in vitro synthesized RNA. Hybridization to unlabeled mercurated cDNA, RNase A digestion, and subsequent purification of the hybrids on SH-agarose allowed the quantitation of newly synthesized ovalbumin mRNA sequences. Approximately 0.1% of the newly synthesized RNA was identified as ovalbumin RNA by both methods. The synthesis of ovalbumin RNA progressed during the incubation of nuclei and was sensitive to actinomycin D and low concentrations of alpha-amanitin. The preferential in vitro transcription of the ovalbumin gene (1000-fold over random transcription of the chicken genome) by RNA polymerase B (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) suggests that the specificity of in vitro RNA synthesis is retained in isolated nuclei.
mesh_terms: D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002645:Chickens; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D003609:Dactinomycin / Q000494:pharmacology; D066298:In Vitro Techniques; D008628:Mercury; D010047:Ovalbumin / Q000096:biosynthesis*; D010057:Oviducts / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis*; D014158:Transcription, Genetic
chemical_list: D000546:Amanitins; D012333:RNA, Messenger; D003609:Dactinomycin; D010047:Ovalbumin; D012321:DNA-Directed RNA Polymerases; D008628:Mercury
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