Sabel'nikov A G, Avdeeva A V, Il'iashenko B I
Genetika. 1977;13(7):1281-8.
Escherichia coli K-12 cells grown to early and late exponential, and early and late stationary phases were treated with CA2+ and analysed for the ability of exogenous 14C-DNA uptake and genetic transformation. DNA-membrane complexes were detected detected by isopicnic centrifugation of cell lysates in sucrose density gradient. It is found that 1) during all the tested phases of the growth cycle, E. coli cells attain the ability of enhanced DNA uptake after Ca2+ treatment; 2) exogenous and host DNAs are associated with the cell membrane during all tested growth phases; 3) nevertheless, the ability to form transformants is strongly time-dependent: the cells can be transformed during logarithmic phase only; 4) Ca2+ protects exogenous DNA against its degradation by bovine pancreatic DNAase. The peculiarities of Ca2+-induced competence, actual and possible interference of Ca2+ at different transformation steps are briefly discussed.
将生长至指数早期、晚期以及稳定期早期、晚期的大肠杆菌K - 12细胞用Ca²⁺处理,并分析其摄取外源¹⁴C - DNA和遗传转化的能力。通过在蔗糖密度梯度中对细胞裂解物进行等密度离心来检测DNA - 膜复合物。结果发现:1)在生长周期的所有测试阶段,经Ca²⁺处理后大肠杆菌细胞均获得增强的DNA摄取能力;2)在所有测试的生长阶段,外源DNA和宿主DNA均与细胞膜相关联;3)然而,形成转化体的能力强烈依赖于时间:细胞仅能在对数期被转化;4)Ca²⁺保护外源DNA不被牛胰DNA酶降解。本文简要讨论了Ca²⁺诱导的感受态的特点,以及Ca²⁺在不同转化步骤中实际的和可能的干扰。