Journal
PLASMID
Volume 64, Issue 3, Pages 170-176Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.plasmid.2010.07.003
Keywords
Heterologous; Integrative; Probiotic; Therapeutic; Recombinant; Replicative plasmid and secretion
Categories
Funding
- CONACyT [82010, 204213]
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Integrative and replicative plasmids for the expression driven by the P-43 promoter and secretion of recombinant proteins in Bacillus subtilis were constructed. The plasmids named pInt and pRep respectively were tested for the production of recombinant human interferon gamma (rhIFN-gamma). A synthetic hIFN-gamma gene employing the optimized B. subtilis codon usage was fused with the Bacillus licheniformis alpha-amylase signal peptide (sp-amyL) encoding sequence. The integrative construct produced 2.5 +/- 0.2 mg l(-1) and the replicative system produced 20.3 +/- 0.8 mg l(-1) of total recombinant rhIFN-gamma. The results showed that secretion of hIFN-gamma was the bottleneck for the overexpression of mature rhIEN-gamma by B. subtilis. (C) 2010 Elsevier Inc. All rights reserved.
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