Use of the human hepcidin gene to build a positive-selection vector for periplasmic expression in Escherichia coli |
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Authors: | Jé rome Haustant,Annesha Sil,Christopher Maillo-Rius,Agnè s Hocquellet,Patricia Costaglioli,Bertrand Garbay,Wilfrid Dieryck |
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Affiliation: | 1. Université de Bordeaux, EA 4135, F-33000 Bordeaux, France;2. Bordeaux-INP, EA4135, F-33000 Bordeaux, France |
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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. |
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Keywords: | Positive selection Hepcidin Expression vector Cloning |
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