Ming P M, Chang H L, Baserga R
Proc Natl Acad Sci U S A. 1976 Jun;73(6):2052-5. doi: 10.1073/pnas.73.6.2052.
A temperature-sensitive mutant of Syrian hamster cells, AF8 was fused with simian-virus-40-transformed Lesch-Nyhan fibroblasts; LNSV, in the presence of beta-propiolactone-inactivated Sendai virus. The AF8 cells grow well at 33.5 degrees but are arrested in mid G1 period when shifted to 39 degrees. The LNSV cells are deficient in hypoxanthine guanine phosphoribosyltransferase. The hybrid clones were selected in hypoxanthine-aminopterin-thymidine medium at 39 degrees. A total of 20 clones was isolated and karyotyped. All clones contained most of the hamster chromosomes and one to eight human chromosomes. The preferential retention of human chromosome 3 was observed in 100% of the metaphases of all clones. In nine of the clones, the only human chromosome present was chromosome 3. The results indicate that human chromosome 3 is responsible for conferring to the hybrid cells the ability to grow at nonpermissive temperature for AF8 cells, i.e., the ability to overcome the G1 block. These findings, together with other reports in the cell cycle may be regulated by genes located on different chromosomes.
叙利亚仓鼠细胞的一个温度敏感突变体AF8与猿猴病毒40转化的莱施-尼汉成纤维细胞LNSV在β-丙内酯灭活的仙台病毒存在下进行融合。AF8细胞在33.5摄氏度时生长良好,但转移到39摄氏度时会在G1期中期停滞。LNSV细胞缺乏次黄嘌呤鸟嘌呤磷酸核糖转移酶。杂交克隆在39摄氏度的次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷培养基中进行筛选。总共分离出20个克隆并进行了核型分析。所有克隆都包含大部分仓鼠染色体和1至8条人类染色体。在所有克隆的100%中期相中都观察到了人类3号染色体的优先保留。在9个克隆中,唯一存在的人类染色体就是3号染色体。结果表明,人类3号染色体赋予杂交细胞在AF8细胞的非允许温度下生长的能力,即克服G1期阻滞的能力。这些发现,连同细胞周期中的其他报道,可能表明细胞周期是由位于不同染色体上的基因调控的。