Bender R A, Sambucetti L C
Mol Gen Genet. 1983;189(2):263-8. doi: 10.1007/BF00337815.
Klebsiella aerogenes recombinants resulting from bacteriophage P1-mediated generalized transduction failed to increase in number for approximately six generations after transduction. Nevertheless these recombinants continued to grow and became sensitive to penicillin after a transient resistance, suggesting that the cells were growing as long, non-dividing filaments. When filamentous cells were isolated from transduced cultures by gradient centrifugation, recombinants were 1000-fold more frequent among the filaments than among the normal-sized cells. The suppression of cell-division lasted for six generations whether markers near the origin (gln, ilv) or terminus (his, trp) of chromosome replication were used, despite a 50-fold difference in transduction frequencies for these markers. The suppression of cell division was a host response to recombination rather than to P1 invasion since cells lysogenized by P1 in these same experiments showed only a short (two generation) suppression of cell division. We speculate that the suppression of cell-division is an SOS response triggered by the degraded DNA not incorporated in the final recombinant. We demonstrate that both the filamentation and the transient penicillin resistance of recombinant cells can be exploited to enrich greatly for recombinants, raising transduction frequencies to as high as 10(-3).
由噬菌体P1介导的普遍性转导产生的产气克雷伯菌重组体在转导后大约六代内数量未能增加。然而,这些重组体继续生长,并在短暂耐药后对青霉素变得敏感,这表明细胞是以长的、不分裂的细丝形式生长。当通过梯度离心从转导培养物中分离出丝状细胞时,重组体在细丝中的出现频率比在正常大小的细胞中高1000倍。无论使用靠近染色体复制起点(谷氨酰胺、异亮氨酸)还是终点(组氨酸、色氨酸)的标记,细胞分裂的抑制都持续了六代,尽管这些标记的转导频率相差50倍。细胞分裂的抑制是宿主对重组的反应,而不是对P1入侵的反应,因为在这些相同实验中被P1溶源化的细胞仅表现出短暂(两代)的细胞分裂抑制。我们推测细胞分裂的抑制是由未整合到最终重组体中的降解DNA触发的SOS反应。我们证明重组细胞的丝化和短暂青霉素耐药性都可用于极大地富集重组体,将转导频率提高到高达10^(-3)。