Luss E V, Volkova L V, Moiseenko E V, Petrova O N, Shapiro N I
Genetika. 1981;17(12):2172-83.
After polyethylene glycol treatment of GPRT- mutant cells, 12 clones were isolated on the ATG medium showing intragenic complementation. Karyological analysis confirmed the hybrid nature of the clones isolated. The GPRT activity in the hybrid clones, as assessed in vitro, exceeded the sum of parental activities. In vivo incorporation of 14C-hypoxanthine showed the GPRT activity in the hybrids to be an order of magnitude higher than in the mutant parental cells. Moreover, the GPRT activity in the hybrid clones was found to increase considerably during cultivation on the ATG medium; hence, their ability to multiply on this selective medium. All hybrid cells surviving and multiplying on the ATG medium were shown to maintain a high enough resistance to 8-AG, 6-MP and, to somewhat less extent, to 6-TG. The frequency of complementation was determined for the cells of mutant clones selected on media with different purine base analogues. The complementation map for the GPRT locus was constructed and proved to be linear. Five groups of complementation were specified.
用聚乙二醇处理GPRT突变细胞后,在显示基因内互补的ATG培养基上分离出12个克隆。染色体分析证实了所分离克隆的杂种性质。体外评估显示,杂种克隆中的GPRT活性超过了亲本活性之和。体内14C-次黄嘌呤掺入显示,杂种中的GPRT活性比突变亲本细胞高一个数量级。此外,发现杂种克隆在ATG培养基上培养期间GPRT活性显著增加;因此,它们能够在这种选择培养基上增殖。所有在ATG培养基上存活并增殖的杂种细胞对8-AG、6-MP以及对6-TG的抗性略低,均表现出足够高的抗性。测定了在含有不同嘌呤碱基类似物的培养基上选择的突变克隆细胞的互补频率。构建了GPRT基因座的互补图谱,证明其为线性。确定了五组互补。