Immunochemical isolation and characterization of vitellogenin mRNA from liver of estradiol-treated chicks.
Vitellogenin mRNA was purified through three steps. A heavy polysome fraction was obtained by discontinuous sucrose density gradient centrifugation, vitellogenin polysomes were immunoprecipitated with affinity-chromatography-purified anti-lipovitellin IgG and goat anti-rabbit IgG, the enriched mRNA was isolated on a poly(U)-Sepharose column. As judged by its specific activity in a reticulocyte lysate system, vitellogenin mRNA has been enriched a 1000-fold with a recovery of 30%. On 99% formamide 3.4% polyacrylamide gels vitellogenin mRNA has an Mr of 2.4-2.5 X 10(6) and codes for a peptide of Mr 240000, which under our incubation conditions is partially degraded to smaller peptides.
D000818:Animals; D002645:Chickens; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D007700:Kinetics; D008074:Lipoproteins / Q000096:biosynthesis*; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism; D011233:Precipitin Tests; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D014819:Vitellogenins / Q000096:biosynthesis*
D008074:Lipoproteins; D012333:RNA, Messenger; D014819:Vitellogenins; D004958:Estradiol