从大鼠肝脏中分离和部分纯化铜蓝蛋白信使核糖核酸

Isolation and partial purification of ceruloplasmin messenger RNA from rat liver.

作者信息

Neifakh S A, Gaitskhoki V S, Klimov N A, Puchkova L V, Shavlovski M M, Schwartzman A L

出版信息

Mol Biol Rep. 1977 Mar;3(3):235-42. doi: 10.1007/BF00643479.

Abstract

Partially purified ceruloplasmin mRNA was isolated using indirect immunoprecipitation of rat liver polysomes and poly(U)-Sepharose chromatography of polysomal RNA. This RNA programmed the synthesis of ceruloplasmin polypeptides in a cell-free system from mitochondria. Immunochemical analysis of the translation products revealed a 40-fold enrichment of the ceruloplasmin mRNA activity. The purified ceruloplasmin mRNA migrated as a major homogeneous component with an apparent molecular weight about 1 X 10(6) daltons in polyacrylamide gels containing sodium dodecyl sulfate. The immunoprecipitated products of the cell-free translation had molecular weights in the range 4.5--5.4 X 10(4) daltons as estimated by gel-electrophoresis under denaturating conditions. These values approach the weight of the half-molecule of native ceruloplasmin.

摘要

利用大鼠肝脏多核糖体的间接免疫沉淀和多核糖体RNA的聚尿苷酸-琼脂糖层析法分离出部分纯化的铜蓝蛋白mRNA。这种RNA在来自线粒体的无细胞系统中指导铜蓝蛋白多肽的合成。对翻译产物的免疫化学分析显示铜蓝蛋白mRNA活性富集了40倍。在含有十二烷基硫酸钠的聚丙烯酰胺凝胶中，纯化的铜蓝蛋白mRNA以主要的均一成分迁移，其表观分子量约为1×10⁶道尔顿。在变性条件下通过凝胶电泳估计，无细胞翻译的免疫沉淀产物的分子量在4.5 - 5.4×10⁴道尔顿范围内。这些值接近天然铜蓝蛋白半分子的重量。