Ikeda H, Kobayashi I
Proc Natl Acad Sci U S A. 1977 Sep;74(9):3932-6. doi: 10.1073/pnas.74.9.3932.
Recombinant DNA molecule of phage lambda formed in Escherichia coli in the presence of chloramphenicol and/or rifampin can be assayed by their biological activity. recA- cells were found to be capable of forming recombinant lambda phage DNA in the presence of chloramphenicol. The relatively high recA-independent recombination observed in this system contrasts with the relatively low recA-independent recombination when recombinant phage particles rather than recombinant DNA are titrated. Formation of the recombinant DNA was suppressed by the the addition of rifampin. The introduction of the rif-r mutation into host bacteria made their recombination activity rifampin-resistant. These results show that DNA-dependent RNA polymerase (EC 2.7.7.6) is involved in this recA-independent pathway of recombination, which is named the "Rpo pathway." This is distinct from Red, Int, RecBC, RecE, or Der pathways of recombination. Crossover was much more frequent in the N-PL-cI and cI-PR-O regions than in the A-D and O-S regions. The crossover seems to occur in the regions that are transcribed actively. Some local change of DNA structure caused by transcription might be required for the Rpo pathway of recombination.
在氯霉素和/或利福平存在的情况下于大肠杆菌中形成的噬菌体λ重组DNA分子,可以通过其生物学活性进行检测。发现recA-细胞能够在氯霉素存在的情况下形成重组λ噬菌体DNA。在该系统中观察到的相对较高的不依赖recA的重组与滴定重组噬菌体颗粒而非重组DNA时相对较低的不依赖recA的重组形成对比。利福平的添加抑制了重组DNA的形成。将rif-r突变引入宿主细菌使其重组活性对利福平具有抗性。这些结果表明,DNA依赖性RNA聚合酶(EC 2.7.7.6)参与了这种不依赖recA的重组途径,该途径被命名为“Rpo途径”。这与Red、Int、RecBC、RecE或Der重组途径不同。在N-PL-cI和cI-PR-O区域的交叉比在A-D和O-S区域更频繁。交叉似乎发生在活跃转录的区域。转录引起的DNA结构的一些局部变化可能是Rpo重组途径所必需的。