Cloning, auto-induction expression, and purification of rSpaA swine erysipelas antigen.

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.

• Authors: Adilson José da Silva, Mônica Rosas da Costa Iemma, Antônio Carlos Luperni Horta, Cíntia Regina Sargo, Raquel de Lima Camargo Giordano, Roberto de Campos Giordano, Teresa Cristina Zangirolami, Maria Teresa Marques Novo
• Journal: Current microbiology (Curr Microbiol) Vol. 65 Issue 4 Pg. 369-74 (Oct 2012) ISSN: 1432-0991 [Electronic] United States
• PMID: 22729211 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Bacterial
• Bacterial Proteins
• Recombinant Proteins
• SpaA protein, Erysipelothrix rhusiopathiae

Topics

• Animals
• Antigens, Bacterial (chemistry, genetics, isolation & purification)
• Bacterial Proteins (chemistry, genetics)
• Chromatography, Affinity
• Cloning, Molecular
• Erysipelothrix (genetics)
• Escherichia coli (genetics)
• Gene Expression
• Inclusion Bodies
• Molecular Weight
• Protein Denaturation
• Recombinant Proteins (chemistry, genetics, isolation & purification)
• Swine
• Swine Erysipelas (immunology, microbiology)