Stanley P, Sallustio S, Krag S S, Dunn B
Department Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461.
Somat Cell Mol Genet. 1990 May;16(3):211-23. doi: 10.1007/BF01233357.
In attempts to isolate new CHO glycosylation mutants, selection protocols using plant lectins that bind galactose residues of cell surface carbohydrates were applied to mutagenized CHO populations. The lectins were used alone or in combination to obtain seven ricin-resistant phenotypes. Each mutant had distinctive properties compared with previously described ricin-resistant CHO cells. One of the new phenotypes was dominant in somatic cell hybrids, and the others were recessive. Complementation analyses between related lectin-resistant (LecR) phenotypes indicated that each new isolate represented a novel genotype. Five of the mutants had properties typical of new CHO glycosylation mutants. The remaining two mutants were not readily categorized. Although they did not appear to be ricin-internalization or protein-synthesis mutants, they also did not display the marked alterations in sensitivity to several lectins of different sugar specificity expected for glycosylation mutants. The seven new LecR mutants described in these studies brings the total number of different LecR CHO mutants isolated by this and other laboratories to about 40. Criteria for identifying new LecR mutations in CHO cells are discussed.
为了分离新的中国仓鼠卵巢细胞（CHO）糖基化突变体，将使用能结合细胞表面碳水化合物半乳糖残基的植物凝集素的筛选方案应用于诱变后的CHO细胞群体。这些凝集素单独使用或联合使用，以获得七种蓖麻毒素抗性表型。与先前描述的蓖麻毒素抗性CHO细胞相比，每个突变体都有独特的特性。其中一种新表型在体细胞杂种中占主导地位，其他则为隐性。相关凝集素抗性（LecR）表型之间的互补分析表明，每个新分离株代表一种新的基因型。其中五个突变体具有新的CHO糖基化突变体的典型特性。其余两个突变体不容易归类。虽然它们似乎不是蓖麻毒素内化或蛋白质合成突变体，但它们也没有表现出糖基化突变体预期的对几种具有不同糖特异性的凝集素敏感性的明显改变。这些研究中描述的七个新的LecR突变体使该实验室和其他实验室分离出的不同LecR CHO突变体总数达到约40个。本文还讨论了鉴定CHO细胞中新的LecR突变的标准。
