Cloning, Auto-induction Expression, and Purification of rSpaA Swine Erysipelas Antigen

被引：8

作者：

da Silva, Adilson Jose ^{[1
]}

da Costa Iemma, Monica Rosas ^{[1
]}

Luperni Horta, Antonio Carlos ^{[1
]}

Sargo, Cintia Regina ^{[1
]}

Camargo Giordano, Raquel de Lima ^{[1
]}

Giordano, Roberto de Campos ^{[1
]}

Zangirolami, Teresa Cristina ^{[1
]}

Marques Novo, Maria Teresa ^{[2
]}

机构：

[1] Univ Fed Sao Carlos, Dept Chem Engn, BR-13565905 Sao Carlos, SP, Brazil

[2] Univ Fed Sao Carlos, Genet & Evolut Dept, BR-13565905 Sao Carlos, SP, Brazil

来源：

CURRENT MICROBIOLOGY | 2012年 / 65卷 / 04期

基金：

巴西圣保罗研究基金会;

关键词：

PROTECTIVE ANTIGEN; RHUSIOPATHIAE; PROTEINS; CHALLENGE; DNA;

D O I：

10.1007/s00284-012-0171-y

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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.

引用

页码：369 / 374

页数：6

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