Sano H, Spaeth E, Burton W G
Eur J Biochem. 1979 Jan 2;93(1):173-80. doi: 10.1111/j.1432-1033.1979.tb12808.x.
Polysomes specifically synthesizing the large subunit of ribulose-1,5-bisphosphate carboxylase were isolated from Chlamydomonas reinhardi cells by the indirect immunoprecipitation method. Electrophoretic analysis showed that the immunoprecipitated polysomes were of chloroplast origin. The mRNA coding for the large subunit which was purified from immunoprecipitated polysomes migrated at the 19-S position on sucrose density gradients, and its molecular weight was estimated to be 7.3 x 10(5) by acid-urea/agarose gel electrophoresis. The mRNA was translated in vivo with a cell-free protein-synthesizing system derived from Escherichia coli to give full-length large-subunit polypeptides.
通过间接免疫沉淀法从莱茵衣藻细胞中分离出特异性合成1,5-二磷酸核酮糖羧化酶大亚基的多核糖体。电泳分析表明，免疫沉淀的多核糖体起源于叶绿体。从免疫沉淀的多核糖体中纯化出的编码大亚基的mRNA在蔗糖密度梯度上迁移到19-S位置，通过酸性尿素/琼脂糖凝胶电泳估计其分子量为7.3×10⁵。该mRNA在体内用源自大肠杆菌的无细胞蛋白质合成系统进行翻译，以产生全长的大亚基多肽。
