Abstract
Summary An E. coli «permeabilized cell system (PCS) is described, resulting from 2 M-sucrose plasmolysis in the presence of 10−4 M EDTA. This treatment induces some interesting properties in the cells: permeability to various phosphorylated derivatives without osmotical fragility, depletion from endogenous metabolites and lack of growth on rich medium. This system appears well suited for the study of uncoupled macromolecular synthesis: o (1) in the presence of the usual RNA polymerase substrates (ATP, CTP, GTP and UTP), PCS prominently synthesizes messenger RNA of exceptional great stability (half-life greater than 60 minutes). The reasons of this stability will be discussed; (2) the mRNAs thus formed are active in protein synthesis as it can be shown by a stepwise induction experiment: RNA synthesized in the presence of the four ribonucleoside triphosphates plus IPTG can direct the synthesis of β-galactosidase, after PCS is transferred in an amino acid medium plus actinomycin D; (3) when PCS is supplemented with a complete amino mixture in addition to the four ribonucleoside triphosphates, it synthesizes proteins and also accumulates large amounts of ribosomal and transfer RNA's.
Published Version
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