HAUSTANT, Jerome; SIL, Annesha; MAILLO-RIUS, Christopher; HOCQUELLET, Agnes; COSTAGLIOLI, Patricia; GARBAY, Bertrand; DIERYCK, Wilfrid

doi:10.1016/j.ab.2016.02.002

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HAUSTANT, Jerome

SIL, Annesha

MAILLO-RIUS, Christopher

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Analytical biochemistry. 2016, vol. 500, p. 35-7

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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. Copyright 2016 Elsevier Inc. All rights reserved.< Leer menos

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Use of the human hepcidin gene to build a positive-selection vector for periplasmic expression in Escherichia coli

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