title: Use of the human hepcidin gene to build a positive-selection vector for periplasmic expression in Escherichia coli.
abstract: Recombinant proteins are often produced in the periplasm of Escherichia coli because this facilitates the purification process. The oxidizing environment favors the formation of disulfide bridges. We showed that the periplasmic expression of the human hormone hepcidin 25 (Hep25) fused to the maltose-binding protein (MBP) resulted in cell death. This toxicity was not observed when MBP-Hep25 accumulated in the bacterial cytoplasm, or when Hep25 was addressed to the periplasm without the MBP tag. We then modified the periplasmic expression vector pMALp2E to create pMALp2EH, a positive-selection vector with Hep25 as counterselection gene.
authors.affiliation: Université de Bordeaux, EA 4135, F-33000 Bordeaux, France; Bordeaux-INP, EA4135, F-33000 Bordeaux, France.
authors.forename: Jérome
authors.initials: J
authors.lastname: Haustant
authors.affiliation: Université de Bordeaux, EA 4135, F-33000 Bordeaux, France; Bordeaux-INP, EA4135, F-33000 Bordeaux, France.
authors.forename: Annesha
authors.initials: A
authors.lastname: Sil
authors.affiliation: Université de Bordeaux, EA 4135, F-33000 Bordeaux, France; Bordeaux-INP, EA4135, F-33000 Bordeaux, France.
authors.forename: Christopher
authors.initials: C
authors.lastname: Maillo-Rius
authors.affiliation: Université de Bordeaux, EA 4135, F-33000 Bordeaux, France; Bordeaux-INP, EA4135, F-33000 Bordeaux, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Hocquellet
authors.affiliation: Université de Bordeaux, EA 4135, F-33000 Bordeaux, France; Bordeaux-INP, EA4135, F-33000 Bordeaux, France.
authors.forename: Patricia
authors.initials: P
authors.lastname: Costaglioli
authors.affiliation: Université de Bordeaux, EA 4135, F-33000 Bordeaux, France; Bordeaux-INP, EA4135, F-33000 Bordeaux, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Garbay
authors.affiliation: Université de Bordeaux, EA 4135, F-33000 Bordeaux, France; Bordeaux-INP, EA4135, F-33000 Bordeaux, France. Electronic address: wilfrid.dieryck@bordeaux-inp.fr.
authors.forename: Wilfrid
authors.initials: W
authors.lastname: Dieryck
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D064451:Hepcidins / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D019897:Periplasm / Q000378:metabolism*
chemical_list: D064451:Hepcidins
keywords: Cloning; Expression vector; Hepcidin; Positive selection