Wullems G J, van der Horst J, Bootsma D
Somatic Cell Genet. 1976 Jul;2(4):359-71. doi: 10.1007/BF01538840.
Evidence is presented for the uptake of the human X chromosome by human-Chinese hamster cell hybrids which lack H P R T activity, following incubation with isolated human HeLa S3 chromosomes. Sixteen independent clonal cell lines were isolated in H A T medium, all of which contained a human X chromosome as determined by trypsin-Giemsa staining. The frequency of H A T-resistant clones was 32 x 10(-6) when 10(7) cells were incubated with 10(8) HeLa chromosomes. Potential reversion of the hybrid cells in H A T medium was less than 5 x 10(-7). The 16 isolated cell lines all contained activity of the human X-linked marker enzymes H P R T, P G K,alpha-Gal A, and G6PD, as determined by electrophoresis. The phenotype of G6PD was G6PD A, corresponding to G6PD A in HeLa cells. The human parental cells used in the fusion to form the hybrids had the G6PD B phenotype. The recipient cells gave no evidence of containing human X chromosomes. These results indicate that incorporation and expression of HeLa X chromosomes is accomplished in human-Chinese hamster hybrids which lack a human X chromosome.
有证据表明,将缺乏次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(HPRT)活性的人 - 中国仓鼠细胞杂种与分离的人HeLa S3染色体一起孵育后,它们摄取了人类X染色体。在次黄嘌呤 - 氨基蝶呤 - 胸腺嘧啶核苷(HAT)培养基中分离出16个独立的克隆细胞系,通过胰蛋白酶 - 吉姆萨染色确定,所有这些细胞系都含有一条人类X染色体。当10⁷个细胞与10⁸条HeLa染色体一起孵育时,HAT抗性克隆的频率为32×10⁻⁶。杂种细胞在HAT培养基中的潜在回复突变率小于5×10⁻⁷。通过电泳测定,16个分离的细胞系均含有人类X连锁标记酶HPRT、磷酸甘油酸激酶(PGK)、α - 半乳糖苷酶A(α - Gal A)和葡萄糖 - 6 - 磷酸脱氢酶(G6PD)的活性。G6PD的表型为G6PD A,与HeLa细胞中的G6PD A相对应。用于融合形成杂种的人类亲代细胞具有G6PD B表型。受体细胞没有证据表明含有人类X染色体。这些结果表明,在缺乏人类X染色体的人 - 中国仓鼠杂种中,HeLa X染色体的掺入和表达得以实现。