Stamato T D, Jones C
Somatic Cell Genet. 1977 Nov;3(6):639-47. doi: 10.1007/BF01539071.
A mutant Chinese hamster ovary cell deficient in lactate dehydrogenase A activity has been isolated using a nonselective technique. The method uses histochemical staining to examine colonies directly for enzyme activity and nylon cloth replica plating to recover particular clones. The mutant cell has an apparent Km (pyruvate to lactate) that is nearly tenfold higher than the parental cell, while its Vmax has been reduced more than 80-fold. In mutant cell extracts with added porcine LDH-B enzyme, molecular dissociation and recombination of subunits produces two new active LDH tetramers (A1B3, A2B2). The electrophoretic mobility of at least one of the tetramers (A1B3) was different from those formed in the parental extracts. The evidence suggests the variant cell contains a mutation in the structural gene for LDH-A.
利用一种非选择性技术分离出了一株乳酸脱氢酶A活性缺陷的中国仓鼠卵巢突变细胞。该方法使用组织化学染色直接检查菌落的酶活性，并通过尼龙布影印平板法回收特定的克隆。突变细胞的表观Km（丙酮酸转化为乳酸）比亲代细胞高近10倍，而其Vmax降低了80倍以上。在添加了猪LDH-B酶的突变细胞提取物中，亚基的分子解离和重组产生了两种新的活性LDH四聚体（A1B3、A2B2）。至少一种四聚体（A1B3）的电泳迁移率与亲代提取物中形成的不同。证据表明该变异细胞在LDH-A的结构基因中存在突变。
