Cloning, auto-induction expression, and purification of rSpaA swine erysipelas antigen.
Curr Microbiol
; 65(4): 369-74, 2012 Oct.
Article
em En
| MEDLINE
| ID: mdl-22729211
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Erisipela Suína
/
Proteínas de Bactérias
/
Erysipelothrix
/
Antígenos de Bactérias
Limite:
Animals
Idioma:
En
Revista:
Curr Microbiol
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Brasil
País de publicação:
Estados Unidos