Mertvetsov N P, Chesnokov V N, Blinova N N, Golovin S Ia, Salganik R I
Mol Biol (Mosk). 1978 Jul-Aug;12(4):806-13.
Messenger RNA (mRNA) capable of coding hydrocortisone induced isoenzyme of tyrosineaminotransferase (TAT) was isolated and purified approximately 5000 times. Highly specific rabbit antibodies to the isoenzyme were obtained. Using these antibodies a specific antibody immunoadsorbent with a high capacity for specific binding to TAT was prepared. From total rat liver polyribosomes induced by hydrocortisone a specific fraction synthesising TAT was isolated by fractionation on the antibody immunoadsorbent. Affinity chromatography using poly(U)--Sephadex 4B allowed to isolate a poly(A)-containing RNA from this polyribosome fraction. The mRNA (15-16S) acts as a matrix in a cell-free protein synthesising system. The extent of purification of the mRNA was estimated by the synthesis of specific protein TAT in the cell-free system.
能够编码氢化可的松诱导的酪氨酸转氨酶（TAT）同工酶的信使核糖核酸（mRNA）被分离并纯化了约5000倍。获得了针对该同工酶的高度特异性兔抗体。使用这些抗体制备了一种对TAT具有高特异性结合能力的特异性抗体免疫吸附剂。通过在抗体免疫吸附剂上进行分级分离，从氢化可的松诱导的大鼠肝脏总多核糖体中分离出合成TAT的特定组分。使用聚（U）-葡聚糖凝胶4B进行亲和层析，可从该多核糖体组分中分离出含聚（A）的RNA。该mRNA（15 - 16S）在无细胞蛋白质合成系统中作为模板。通过在无细胞系统中合成特异性蛋白质TAT来估计mRNA的纯化程度。
