Siciliano M J, Siciliano J, Humphrey R M
Proc Natl Acad Sci U S A. 1978 Apr;75(4):1919-23. doi: 10.1073/pnas.75.4.1919.
Electrophoretic shift mutants induced in Chinese hamster ovary (CHO) cells indicate that these cells not extensively functionally hemizygotic. Therefore, effective haploidy is unsatisfactory as a general theory to explain the frequency of recessive mutants in this cell line. CHO cells were screened for electrophoretic shift variants of enzymes coded by approximately 40 genetic loci. Clones isolated after exposure to ultraviolet radiation were examined by starch gel and Cellogel electrophoresis. Shift variants were recovered for enzymes representing 11 different loci. Variant clones were subcloned to demonstrate the heritability of the variations Mutants at nine loci produced multiple-banded patterns consistent with the patterns expected of genes at loci represented twice (diploid). Chromosome localization of these diploid loci in other mammalian species where they have been mapped, suggests that they represent a random sample of CHO genes. Chromosome analysis of mutant subclones indicated that the variation did not take place in tetraploid cells. The data indicate that the quasi-diploid CHO cells appear only as functionally hemizygous as would be expected of a slightly hypodiploid cell line derived from an organism in which the haploid number is 11.
在中国仓鼠卵巢(CHO)细胞中诱导产生的电泳迁移突变体表明,这些细胞在功能上并非广泛半合子状态。因此,有效的单倍体状态作为一种普遍理论,用以解释该细胞系中隐性突变体的频率并不令人满意。对CHO细胞进行筛选,以寻找由大约40个基因位点编码的酶的电泳迁移变体。经紫外线照射后分离得到的克隆,通过淀粉凝胶电泳和Cellogel电泳进行检测。回收了代表11个不同位点的酶的迁移变体。对变体克隆进行亚克隆,以证明变异的遗传性。九个位点的突变体产生了多条带模式,与在两倍体(二倍体)位点上的基因预期模式一致。这些二倍体位点在已绘制图谱的其他哺乳动物物种中的染色体定位表明,它们代表了CHO基因的随机样本。突变亚克隆的染色体分析表明,变异并非发生在四倍体细胞中。数据表明,准二倍体CHO细胞在功能上仅表现出与源自单倍体数为11的生物体的轻度亚二倍体细胞系预期的半合子状态相同。