Cloning, Auto-induction Expression, and Purification of rSpaA Swine Erysipelas Antigen |
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Authors: | da Silva Adilson José da Costa Iemma Mônica Rosas Luperni Horta Antônio Carlos Sargo Cíntia Regina de Lima Camargo Giordano Raquel de Campos Giordano Roberto Zangirolami Teresa Cristina Marques Novo Maria Teresa |
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Institution: | Chemical Engineering Department, Federal University of S?o Carlos, Rodovia Washington Luís, km 235, CP 676, S?o Carlos, SP, CEP 13565-905, Brazil, adiljs@gmail.com. |
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Abstract: | This work reports the cloning, expression, and purification of a 42-kDa fragment of the SpaA protein from Erysipelothrix rhusiopathiae, the main antigenic candidate for a subunit vaccine against swine erysipelas. The use of an auto-induction protocol to improve heterologous protein expression in recombinant Escherichia coli cultures was also investigated. The cellular growth pattern and metabolite formation were evaluated under different induction conditions. The His-tagged protein was over-expressed as inclusion bodies, and was purified by a single chromatography step under denaturing conditions. Auto-induction conditions were shown to be an excellent process strategy, leading to a high level of rSpaA expression (about 25?% of total cellular protein content) in a short period of time. |
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