Denney R M, Craig I W
Biochem Genet. 1976 Feb;14(1-2):99-117. doi: 10.1007/BF00484877.
We describe a simple method for locating tryptophanyl-tRNA synthetase (E.C. 6.1.1.2) on cellulose acetate gels (Cellogel) following electrophoresis. Employing electrophoretic conditions which result in the separation of mouse and human tryptophanyl-tRNA synthetase, we have analyzed extracts of a number of independently derived mouse-human somatic cell hybrids and subclones derived from these hybrids for the presence of human tryptophanyl-tRNA synthetase. Electrophoretic patterns of hybrid extracts which contain human tryptophanyl-tRNA synthetase exhibit three bands. This is consistent with published evidence that the enzyme for mammalian cells is a homologous dimer. The electrophoretic patterns derived from some hybrids are unusual in that the human and hybrid bands of activity are more intense than the mouse band from the same hybrid. An analysis of hybrid cells and extracts indicates that human tryptophanyl-tRNA synthetase segregates with human chromosome 14 and with the only enzyme marker which has previously been assigned to this chromosome, nucleoside phosphorylase.
我们描述了一种在电泳后于醋酸纤维素凝胶(Cellogel)上定位色氨酰 - tRNA合成酶(E.C. 6.1.1.2)的简单方法。利用能分离小鼠和人类色氨酰 - tRNA合成酶的电泳条件,我们分析了许多独立衍生的小鼠 - 人类体细胞杂种以及从这些杂种衍生的亚克隆提取物中人类色氨酰 - tRNA合成酶的存在情况。含有人类色氨酰 - tRNA合成酶的杂种提取物的电泳图谱显示有三条带。这与已发表的证据一致,即哺乳动物细胞的该酶是同源二聚体。一些杂种的电泳图谱不同寻常,在于同一杂种中人类和杂种活性带比小鼠带更强。对杂种细胞和提取物的分析表明,人类色氨酰 - tRNA合成酶与人类14号染色体以及先前唯一被定位到该染色体的酶标记——核苷磷酸化酶一起分离。