Lysis of Escherichia coli with a neutral detergent

Abstract

1. When Escherichia coli cells are treated with lysozyme and versene for 30–45 sec in an ice-bath, they become susceptible to lysis by the neutral detergent Brij-58. 2. The rate of lysis is dependent upon the concentration of the detergent. Brij-58 is approx. 1000-fold less active in lysis of the lysozyme-versene-treated cells than is deoxycholic acid or sodium docecylsulphate. 3. The extent of cell disruption is dependent upon the Mg^(2+) concentration and the ionic strength at the time of lysis. In 70 mM Mg^(2+), only the low molecular weight soluble RNA and soluble protein are released from the cell. In 40 mM Mg^(2+), the 70-S monosomes and the ribosome subunits are released together with the soluble material. At still lower Mg^(2+) concentrations, the polyribosomes are released. 4. Unless the Mg^(2+) concentration is reduced below 5 mM, the cellular DNA, after lysis, may be almost completely sedimented by centrifugation at 3000 × g for 5 min. 5. All strains of E. coli tested were susceptible to lysis by this method.

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