Importance

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Abstract

A method for recombinantly expressing a macromolecule in a host cell is disclosed which involves culturing a host cell which contains two nucleic acid sequences, the first encoding a membrane-permeabilizing agent and the second encoding a desired macromolecule under the operative control of an inducible promoter, to a selected cell density that permits accumulation of the agent. Thereafter, the host cell is exposed to environmental conditions that induce the agent to disrupt the integrity of the cell membrane without complete lysis of the cell membrane, which cancels the energized membrane function. The host cell allows transport through the membrane of small molecular weight compounds. These resulting host cells are cultured in the presence of a nutrient cocktail that contains components that can transport through the disrupted cell membrane, e.g., an inducing agent that induces the tightly regulated promoter and metabolic requirements that permit expression of the macromolecule.

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First Claim