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-γ). A synthetic hIFN-γ gene employing the optimized B. subtilis codon usage was fused with the Bacillus licheniformis α-amylase signal peptide (sp-amyL) encoding sequence. The integrative construct produced 2.5±0.2mgl(-1) and the replicative system produced 20.3±0.8mgl(-1) of total recombinant rhIFN-γ. The results showed that secretion of hIFN-γ was the bottleneck for the overexpression of mature rhIFN-γ by B. subtilis.
Rojas Contreras
Juan A
JA
División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, Lomas 4ª. Sección, San Luis Potosí, SLP, Mexico.
Pedraza-Reyes
Mario
M
Ordoñez
Leandro G
LG
Estrada
Norma Urtiz
NU
Barba de la Rosa
Ana P
AP
De León-Rodríguez
Antonio
A
D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004261:DNA Replication / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005813:Genes, Synthetic; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins; D016415:Sequence Alignment
D011994:Recombinant Proteins; D007371:Interferon-gamma; D004247:DNA