Rudchenko O N, Likhacheva N A, Timakova N V, Il'iashenko B N
Genetika. 1976;12(8):110-5.
The paper is initiated in the aim to use the effect of freezing-thawing in the system of intact Escherichia coli cells treated with Ca++ ions to increase the transfection efficiency. It is demonstrated that freezing-thawing of Ca++ treated cells increases the transfection indices of lambda phage DNA for E. coli Hfr clone with low transfection indices in 20-30 times, and for the strain of E. coli X7026 of high competence-in 5 times. Freezing-thawing efficiently increased the transfection only in cells with a decreasing competence. E. coli Hfr H cells acquired the capacity of more efficient lambda phage DNA perception independently on the growth phase, while in E. coli X7026 cells the maximal transfection increase under the effect of complementary interactions was observed at the beginning of the logariphmic and stationary growth stages, but not during the competence period. Short-term changes emerging in cells during freezing-thawing can probably promote the better penetration of Ca++ ions to active sites on the cell surface, which produces favourable conditions for lambda phage DNA formation.
本文旨在利用冻融作用于经钙离子处理的完整大肠杆菌细胞体系,以提高转染效率。结果表明,冻融处理钙离子处理过的细胞,可使转染指数低的大肠杆菌Hfr克隆对λ噬菌体DNA的转染指数提高20至30倍,而对于感受态高的大肠杆菌X7026菌株,转染指数提高5倍。冻融仅在感受态降低的细胞中有效提高转染效率。大肠杆菌Hfr H细胞在不同生长阶段均获得了更高效摄取λ噬菌体DNA的能力,而在大肠杆菌X7026细胞中,在对数生长期和稳定期开始时,互补相互作用的影响下转染增加最大,而在感受态时期则未观察到。冻融过程中细胞出现的短期变化可能促进钙离子更好地渗透到细胞表面的活性位点,从而为λ噬菌体DNA的形成创造有利条件。