Open AccessSynthesis of Albumin and Malic Enzyme in Wheat‐Germ Lysates and Xenopus laevis Oocytes Programmed with Chicken‐Liver Messenger RNA
Author(s) -
GOODRIDGE Alan,
CIVELLI Olivier,
SCHERRER Klaus,
YIP Cecil
Publication year - 1979
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1979.tb13006.x
Subject(s) - xenopus , messenger rna , biology , rna , microbiology and biotechnology , biochemistry , ribonuclease , polyadenylation , enzyme , protein biosynthesis , centrifugation , homogenization (climate) , gene , biodiversity , ecology
Undegraded, biologically‐active, polyadenylated RNA was isolated from chicken liver by a rapid, simple procedure. Liver cells were dispersed mechanically and then broken gently by controlled Dounce homogenization in the absence of detergent or ribonuclease inhibitors. After removing lysosomes and mitochondria by centrifugation, RNA was precipitated at pH 5.2. Polyadenylated mRNA was isolated directly from the detergent‐solubilized precipitate by oligo(dT)‐cellulose chromatography. The resulting RNA was translated into liver‐specific peptides in both the wheat germ lysate and Xenopus laevis oocytes. Translatable albumin mRNA was detected in the liver cytoplasm of both fed 3‐week‐old chicks and unfed, day‐old chicks. Translatable malic enzyme mRNA was only detected in the livers from the fed chicks.