Gorodetskiĭ S I, Sulimova G E, Popendikite V Kh, Mil'shina N V, Ivanov V N
Biokhimiia. 1984 Apr;49(4):633-43.
Total cellular poly(A+)-RNA was isolated from a lactating cow mammary gland. The poly(A+)-RNA molecules exhibit a heterogeneous distribution from 500 to 5000 nucleotides (average size--1600 nucleotides) and are made up of three main fractions (1550, 950 and 600 nucleotides) possessing a high template activity during translation in vitro. Optimal conditions for poly(A+)-RNA translation in a cell-free protein-synthesizing system from wheat embryos were elaborated. Immunochemical analysis of translation products revealed that 30% of the synthesized polypeptides are precipitated by immunoglobulins against cow milk proteins. Using hybridization with homologous cDNA, the kinetic complexity and heterogeneity of total cellular poly(A+)-RNA were investigated. This population was shown to consist of four classes differing in the diversity of their nucleotide sequences and the number of copies per cell. The total amount of the poly(A+)-RNA species in the cells of a lactating cow mammary gland is 9200, i.e., 0.46% of the genome complexity.
从泌乳奶牛的乳腺中分离出总细胞聚腺苷酸加尾RNA(poly(A+)-RNA)。聚腺苷酸加尾RNA分子呈现出500至5000个核苷酸的异质分布(平均大小为1600个核苷酸),并且由三个主要部分(1550、950和600个核苷酸)组成,这些部分在体外翻译过程中具有高模板活性。阐述了在小麦胚无细胞蛋白质合成系统中聚腺苷酸加尾RNA翻译的最佳条件。对翻译产物的免疫化学分析表明,30%的合成多肽可被抗牛奶蛋白的免疫球蛋白沉淀。利用与同源cDNA的杂交,研究了总细胞聚腺苷酸加尾RNA的动力学复杂性和异质性。该群体显示由四类组成,它们在核苷酸序列的多样性和每个细胞中的拷贝数方面存在差异。泌乳奶牛乳腺细胞中聚腺苷酸加尾RNA种类的总量为9200种,即基因组复杂性的0.46%。