Miller C A, Cohen S N
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1265-9. doi: 10.1073/pnas.75.3.1265.
An endonuclease having EcoRI specificity is produced by bacteria containing the ColE1 plasmid. Such bacterial cells fail to express restriction or modification functions in vivo, and phage or plasmid DNA obtained from ColE1-containing cells has unmodified EcoRI sites that are cleaved in vitro by purified EcoRI endonuclease or by enzyme extracted from bacteria that carry ColE1. No EcoRI DNA methylase activity associated with ColE1 has been detected. The finding of phenotypically cryptic ColE1-dependent EcoRI endonuclease activity and the absence of any detectable EcoRI modification system in ColE1-containing cells suggest a control mechanism that appears to prevent functional expression of the ColE1-determined enzyme in vivo.
具有EcoRI特异性的核酸内切酶由含有ColE1质粒的细菌产生。这类细菌细胞在体内无法表达限制或修饰功能,并且从含ColE1的细胞中获得的噬菌体或质粒DNA具有未修饰的EcoRI位点,这些位点在体外可被纯化的EcoRI核酸内切酶或从携带ColE1的细菌中提取的酶切割。未检测到与ColE1相关的EcoRI DNA甲基化酶活性。在含ColE1的细胞中发现表型隐匿的ColE1依赖性EcoRI核酸内切酶活性,且未检测到任何可检测的EcoRI修饰系统,这表明存在一种控制机制,该机制似乎可防止ColE1决定的酶在体内功能性表达。