Replicative and integrative plasmids for production of human interferon gamma in Bacillus subtilis

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.