Isolation and Characterization of the Messenger RNAs for Mouse Embryonic Globin Chains

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Abstract
MRNA for mouse embryonic globins was purified from yolk sac erythroid cells by oligodeoxythymidilate-cellulose chromatography and sucrose density centrifugation. Full-sized complementary [c] DNA copies of embryonic globin mRNA were synthesized. Acrylamide gel electrophoresis of RNA indicated an average MW of 220,000, including a polyadenylated sequence of about 35 residues, as determined by hybridization to [3H]polyuridylate. Wheat-germ translation products of mRNA were the size of and had the ionic characteristics of the 4 embryonic globin chains .alpha., x, y and z. Hybridization kinetics in vast RNA excess were performed and compared to standard r0t [renaturation time] curves of adult globin messengers, demonstrating a total base sequence complexity of about 880,000 daltons, i.e., 4 different RNA sequences of 220,000 MW. Titration of embryonic globin cDNA with increasing amounts of their complementary RNA templates indicated that embryonic globin messengers were isolated at a high degree of purity.