Varshaver N B, Chernikov V G, Gorbunova L V
Genetika. 1976;12(3):56-65.
Spontaneous and nitrosoguanidine (NG)-induced rate of reversions to glutamine independence was studied in cultured temperature-sensitive glutamine auxotrophs of Chinese hamster cells. In 3 experiments the spontaneous rate of reversions varied from 0.8-10(-6) to 3.84-10(-6) per cell per generation. A dependence of the yield of NG-induced back mutations upon the time interval between the mutagenic treatment and the transfer to selective conditions (glutamine deficient medium, 40 degrees C) was established. No induced revertants were detected when cells were transferred to selective conditions immediately after the treatment with NG. After 2--3 days cultivation in glutamine containing medium at 36 degrees C and the sunsequent transfer to selective conditions the frequency of induced reversions varied from 0.56-10(-4) to 10.55-10(-4) in different experiments; after 6 days -- from 0.05-10(-4) to 4.0-10(-4). In all cases where induction was detected, the difference, between the frequency of glutamine prototrophs in treated and control plates was significant. Glutamine independence proved to be stable after prolonged cultivation under non-selective conditions, the degree of prototrophy being greatly unequal in different clones. No differnce in this respect was detected between spontaneous and NG-induced revertants. The proposed system of reverse mutations can be used for studying diverse problems of somatic cell genetics.
在中国仓鼠细胞的培养温度敏感型谷氨酰胺营养缺陷型中,研究了自发及亚硝基胍(NG)诱导回复为谷氨酰胺非依赖型的速率。在3个实验中,每代每个细胞的自发回复速率在0.8×10⁻⁶至3.84×10⁻⁶之间变化。确定了NG诱导的回复突变产量对诱变处理与转移至选择条件(谷氨酰胺缺乏培养基,40℃)之间时间间隔的依赖性。在用NG处理后立即将细胞转移至选择条件下时,未检测到诱导的回复子。在36℃含谷氨酰胺的培养基中培养2 - 3天,随后转移至选择条件下,不同实验中诱导回复的频率在0.56×10⁻⁴至10.55×10⁻⁴之间变化;培养6天后,频率在0.05×10⁻⁴至4.0×10⁻⁴之间。在所有检测到诱导的情况下,处理平板和对照平板中谷氨酰胺原养型频率之间的差异是显著的。在非选择条件下长时间培养后,谷氨酰胺非依赖型被证明是稳定的,不同克隆中原养型的程度差异很大。在这方面,自发和NG诱导的回复子之间未检测到差异。所提出的回复突变系统可用于研究体细胞遗传学的各种问题。