A mild method for the isolation of folded chromosomes fromEscherichia coli

Abstract

Until now it has not been possible to obtain nuclear bodies from Escherichia coli after treatment with rifampicin. It was generally assumed that the cross-connections between the DNA double strands which are sensitive towards ribonuclease are destroyed under the influence of inhibitors of RNA synthesis like rifampicin. In this paper a new lysis procedure is described for preparing nuclear bodies from E. coli. These particles differ in some respects, especially in salt sensitivity from those prepared by earlier methods. Using the new lysis method it is also possible to obtain folded chromosomes from cells after treatment with rifampicin. These nuclear bodies can be destroyed by ribonuclease. Therefore, it has to be postulated that a fraction of RNA being sufficient to hold the chromosome in the folded shape is not susceptible to the action of rifampicin.