Open AccessImmunochemical Isolation and Characterization of Vitellogenin mRNA from Liver of Estradiol‐Treated Chicks
Author(s) -
JOST JeanPierre,
PEHLING Gundula
Publication year - 1976
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1976.tb10523.x
Subject(s) - vitellogenin , polysome , messenger rna , reticulocyte , microbiology and biotechnology , biology , affinity chromatography , sepharose , trypsinization , polyacrylamide gel electrophoresis , chromatography , biochemistry , incubation , centrifugation , chemistry , rna , trypsin , ribosome , enzyme , gene
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 × 106 and codes for a peptide of M r 240000, which under our incubation conditions is partially degraded to smaller peptides.