title: In vitro RNA synthesis and expression of vitellogenin gene in isolated chicken liver nuclei.
abstract: Optimal conditions for prolonged in vitro synthesis of RNA in isolated chicken liver nuclei have been described. It is shown by incorporation of gamma32P-GTP into RNA, analysis of the product on sucrose density gradient, and digestion with alkaline phosphatase and ribonuclease A that there is reinitiation of RNA synthesis. Polynucleotide kinase activity has been ruled out as explanation for the incorporation of gamma32P-GTP. alpha-Amanitin inhibits RNA synthesis by about 50%. Nuclei prepared from estradiol-treated chicks have twice the RNA synthesis activity as the controls. RNA is synthesized in the presence of Hg-UTP and the mercurated product separated by affinity chromatography on sulfhydryl-Sepharose column under stringent conditions. Vitellogenin mRNA sequences are measured by hybridization with DNA complementary to vitellogenin mRNA. Estradiol treatment leads to a 10-fold increase in vitellogenin mRNA sequences.
mesh_terms: D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002645:Chickens; D004958:Estradiol / Q000494:pharmacology; D005796:Genes; D006160:Guanosine Triphosphate / Q000378:metabolism; D066298:In Vitro Techniques; D008074:Lipoproteins / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008628:Mercury / Q000494:pharmacology; D012313:RNA / Q000096:biosynthesis*; D014544:Uridine Triphosphate / Q000494:pharmacology; D014819:Vitellogenins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000546:Amanitins; D008074:Lipoproteins; D014819:Vitellogenins; D004958:Estradiol; D012313:RNA; D006160:Guanosine Triphosphate; D008628:Mercury; D014544:Uridine Triphosphate