title: [Extracellular production of recombinant human epidermal growth factor (hEGF) in Escherichia coli cells].
abstract: An expression plasmid pPTK-hEGF2 was constructed to provide for the extracellular production of recombinant human epidermal growth factor by the Escherichia coli cells. The plasmid contained two expression cassettes, one of which carried a tandem of the fused genes ompF-hegf under the control of the tac promoter, ensuring regulated secretion of hEGF into the E. coli periplasm, and another one contained the kil gene from the ColE1 plasmid under the control of lac promoter. The regulated low-level biosynthesis of Kil protein increased the permeability of E. coli outer membrane for periplasmic proteins. This enabled the recombinant proteins secreted into the cell periplasm to outflow into the cultural medium. As a result, the E. coli strains that harboured this plasmid construct produced effectively the recombinant hEGF into the cultural medium. The yields of hEGF produced by the nTG1(pPTK-hEGF2) and HB101(pPTK-hEGF2) strains reached 25 and 30 mg/l of cell culture after 14 and 18 h of cultivation, respectively. The hEGF preparation isolated possessed biological activity both in vivo and in vitro.
authors.affiliation: RF GosNII GENETIKA State Research Center, Moscow, Russia. mashko@vnigen.msk.su
authors.forename: L R
authors.initials: LR
authors.lastname: Ptitsyn
authors.forename: I B
authors.initials: IB
authors.lastname: Al'tman
mesh_terms: D003470:Culture Media; D004815:Epidermal Growth Factor / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D005110:Extracellular Space / Q000378:metabolism; D006801:Humans; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor