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The isolation of the plasma membrane from a gram-negative organism.

Abstract A simple and rapid procedure is described for the isolation of plasma membrane from the gram-negative bacterium Rhizobium meliloti. The method involves the mechanical disruption of cells followed by differential and density gradient centrifugation. Cell wall, intermediate, and cell membrane fractions were isolated. Electron microscopy indicated no attachment of the membrane to the wall although some membrane material was trapped within the wall fragments. The intermediate fraction consisted of both wall and membrane material and the membrane fraction appeared to be free of wall material. Biochemical data strongly supported the electron microscopic evidence. Chemical analyses of the uncontaminated membranes indicated that they possessed a composition consistent with other bacterial membranes.