Ullu E, Gambari R, Farace M G, Lunadei M, Fantoni A
Eur J Biochem. 1978 Jul 17;88(1):315-22. doi: 10.1111/j.1432-1033.1978.tb12452.x.
Messenger RNAs for mouse embryonic globins were purified from yolk sac erythroid cells by oligodeoxythymidilate-cellulose chromatography and sucrose density centrifugation. Full-sized complementary DNA copies of embryonic globin mRNAs were synthesized. Acrylamide gel electrophoresis of these RNAs indicate an average molecular weight of 220 000, including a polyadenylated sequence of about 35 residues, as determined by hybridization to [3H]polyuridylate. The wheat-germ translation products of mRNAs have the size and the ionic characteristics of the four embryonic globin chains alpha, x, y and z. Hybridization kinetics in vast RNA excess were performed and compared to standard r0t curves of adult globin messengers, demonstrating a total base sequence complexity of about 880 000 daltons, that is four different RNA sequences of 220 000 molecular weight. The titration of embryonic globin cDNAs with increasing amounts of their complementary RNA templates indicates that the embryonic globin messengers were isolated at a high degree of purity.
通过寡聚脱氧胸苷纤维素层析和蔗糖密度离心从卵黄囊红细胞中纯化出小鼠胚胎珠蛋白的信使核糖核酸（mRNA）。合成了胚胎珠蛋白mRNA的全长互补DNA拷贝。这些RNA的丙烯酰胺凝胶电泳显示其平均分子量为220,000，通过与[3H]聚尿苷酸杂交确定其中包含约35个残基的聚腺苷酸化序列。mRNA的小麦胚芽翻译产物具有α、x、y和z这四条胚胎珠蛋白链的大小和离子特性。在大量RNA过量的情况下进行杂交动力学研究，并与成人珠蛋白信使的标准r0t曲线进行比较，结果表明总碱基序列复杂度约为880,000道尔顿，即四种分子量为220,000的不同RNA序列。用越来越多的互补RNA模板滴定胚胎珠蛋白cDNA表明，所分离的胚胎珠蛋白信使具有高度纯度。
