MESSENGER RNA CORRESPONDING TO MACROPHAGE-ACTIVATION FACTOR, AND ITS PREPARATION

摘要

PURPOSE:To prepare novel mRNA corresponding to macrophage-activation factor (MAF), by selecting MAF-producing cell, culturing the selected cell, and separating the objective mRNA from the cell. CONSTITUTION:Human T-hybridoma obtained by the fusion of human T-lymphocyte and human T-lymphoma cell is used as the MAF-producing cell. The human T-hybrid is cultured, whole RNA is extracted from the cell, and the objective mRNA corresponding to MAF is separated from the whole RNA. The separation is carried out preferably by oligo-dT-cellulose column chromatography or sucrose density-gradient centrifugation. The separated mRNA is translated to protein, and its MAF-activity is determined to identify the mRNA corresponding to MAF. The identified mRNA has the following characteristics. (1) Precipitation constant, 10-12s; (2) polyadenylic acid structure at the 3'-terminal; (3) coding MAF having an isoelectric point of 3 and 5.