Szyf M, Avraham-Haetzni K, Reifman A, Shlomai J, Kaplan F, Oppenheim A, Razin A
Proc Natl Acad Sci U S A. 1984 Jun;81(11):3278-82. doi: 10.1073/pnas.81.11.3278.
Extrachromosomal plasmid DNA is transiently undermethylated in Escherichia coli during amplification in the presence of chloramphenicol. In addition, undermethylation of phage lambda DNA was observed after thermal induction of a lambda c1857 lysogen while the integrated lambda phage DNA was found to be fully methylated. These methylation pattern changes occur under conditions (extensive replication) in which the intracellular methylase level becomes limiting. In an E. coli strain that harbors a plasmid that carries the dam methylase gene and therefore overproduces dam methylase, there is no undermethylation of dam sites in either of the extrachromosomal DNAs. The sites that are methylated by the mec methylase in both plasmid and lambda phage DNAs were undermethylated in the dam overproducer as well. These results indicate that the intracellular level of the E. coli methylase determines the DNA methylation pattern.
在氯霉素存在的情况下进行扩增时,大肠杆菌中的染色体外质粒DNA会短暂地发生低甲基化。此外,热诱导λc1857溶原菌后观察到噬菌体λDNA的低甲基化,而整合的λ噬菌体DNA则被发现是完全甲基化的。这些甲基化模式的变化发生在细胞内甲基化酶水平变得有限的条件下(广泛复制)。在一个携带dam甲基化酶基因的质粒的大肠杆菌菌株中,该菌株因此过量产生dam甲基化酶,染色体外的两种DNA中的dam位点都没有发生低甲基化。在dam甲基化酶过量产生菌中,质粒和λ噬菌体DNA中被mec甲基化酶甲基化的位点也发生了低甲基化。这些结果表明,大肠杆菌甲基化酶的细胞内水平决定了DNA甲基化模式。