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Replicative and integrative plasmids for production of human interferon gamma in Bacillus subtilis |
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| Authors: | Juan A Rojas Contreras Mario Pedraza-Reyes Leandro G Ordoez Norma Urtiz Estrada Ana P Barba de la Rosa Antonio De Len-Rodríguez |
| Institution: | a División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, Lomas 4ª. Sección, C. P. 78216, San Luis Potosí, SLP, Mexico;b Depto. de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, AP 187. Noria Alta S/N, Guanajuato, 36050 Guanajuato, Gto, Mexico;c Facultad de Ciencias Químicas, Universidad Juárez del Estado de Durango, Av. Veterinaria S/N, Col. Valle del Sur, Circuito Universitario, C.P. 34120 Durango, Dgo, Mexico |
| Abstract: | Integrative and replicative plasmids for the expression driven by the P43 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 ± 0.2 mg l−1 and the replicative system produced 20.3 ± 0.8 mg l−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. |
| Keywords: | Heterologous Integrative Probiotic Therapeutic Recombinant Replicative plasmid and secretion |
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