A Method for Extracellular Production of Target Proteins Employing OmpF in E. coli

摘要

PURPOSE: A process for producing a target protein from Escherichia coli through the extracellular secretion using an OmpF gene is provided, therefor the target protein can be simply and massively produced from Escherichia coli through the high concentration of cell cultivation. CONSTITUTION: The process for producing a target protein from Escherichia coli through the extracellular secretion using the OmpF gene comprises the steps of: (i) cloning an oligopeptide expressing a protease-recognition and digestion site and a gene encoding a target protein into an expression vector pOmpF6 containing an ampicillin-resistant gene, an OmpF promoter and the OmpF gene to construct a recombinant expression vector; (ii) transforming a host cell without the OmpF gene with the recombinant expression vector to prepare a transformed cell; (iii) culturing the transformed cell to obtain a OmpF-fusion protein from the cultivated medium; and (iv) treating the OmpF-fusion protein with protease to recover the target protein, wherein the protease is factor Xa, enterokinase, genase, IgA protease, intein, thrombin, trypsin, pepsin, subtilisin or plasmin.