Cloning, auto-induction expression, and purification of rSpaA swine erysipelas antigen.
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.
Chemical Engineering Department, Federal University of São Carlos, Rodovia Washington Luís, km 235, CP 676, São Carlos, SP CEP 13565-905, Brazil. adiljs@gmail.com
Adilson José
AJ
da Silva
Mônica Rosas
MR
da Costa Iemma
Antônio Carlos
AC
Luperni Horta
Cíntia Regina
CR
Sargo
Raquel
R
de Lima Camargo Giordano
Roberto
R
de Campos Giordano
Teresa Cristina
TC
Zangirolami
Maria Teresa
MT
Marques Novo
D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004888:Erysipelothrix / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D002479:Inclusion Bodies; D008970:Molecular Weight; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013552:Swine; D013554:Swine Erysipelas / Q000276:immunology / Q000382:microbiology*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins; C114457:SpaA protein, Erysipelothrix rhusiopathiae