Electron microscopy of membrane-free folded chromosomes from Escherichia coli

Abstract

Membrane-free folded chromosomes were purified from log-phase cultures of Escherichia coli and prepared for electron microscopy by aqueous (Kleinschmidt and Zahn) spreading. The appearance of the chromosomes depended on the salt concentrations in spreading. At certain salt concentration, the chromosomes resembled rosettes, with supercoiled loops of DNA radiating from a central core containing RNA. The rosettes support previous models deduced from physical studies of folded chromosomes. Apparently, cores contain must of the visible RNA, and the organization of the core is linked to the organization of the DNA loops.