Periplasmic expression and recovery of human interferon gamma in Escherichia coli
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A synthetic human interferon gamma (hIFN-γ) gene was fused to SP1 and SP3, two Sec-dependent artificial signal peptides to transport the hIFN-γ to the periplasm of Escherichia coli BL21-SI. The processing efficiency of both SP1-hIFN-γ and SP3-hIFN-γ was dependent on the culture medium as well as the post-induction temperature. Both precursors were processed completely when cells were cultured using minimal medium and a post-induction temperature of 32.5 °C, and only the processed hIFN-γ was detected. The SP3 signal peptide was more efficient than SP1 for the secretion of hIFN-γ. Sixty percent of the total hIFN-γ was secreted to the periplasm using the SP3 signal peptide and a post-induction temperature of 20 °C. Using Tris-sucrose-dithiothreitol (TSD) hypertonic buffer, the periplasmic soluble hINF-γ was recovered with a purity of 85%25. © 2008 Elsevier Inc. All rights reserved.
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Escherichia coli BL21-SI; Osmotic shock; Periplasmic expression; Post-induction temperature; Sec-pathway; Synthetic gene gamma interferon; signal peptide; amino acid sequence; article; biosynthesis; cytoplasm; Escherichia coli; genetics; human; isolation and purification; metabolism; molecular genetics; nucleotide sequence; temperature; ultrastructure; Amino Acid Sequence; Base Sequence; Escherichia coli; Humans; Interferon Type II; Molecular Sequence Data; Periplasm; Protein Sorting Signals; Temperature; Escherichia coli
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