Coexpression of interleukin-6 and -2 from giant panda in Escherichia coli and the biological activity of the fusion protein.
Genet Mol Res
; 12(2): 1987-95, 2013 Jun 14.
Article
em En
| MEDLINE
| ID: mdl-23913382
ABSTRACT
To construct a fusion cytokine protein with more and stronger bioactivities to enhance the immunity of the cytokine alone, we expressed interleukin (IL)-6/(IL)-2 from giant panda (Ailuropoda melanoleuca) in Escherichia coli as a 59.4-kDa fusion protein. Subsequently, the inclusion bodies were solubilized with 8 M urea and applied onto a Ni-nitrilotriacetic acid column. The final production of IL-6/IL-2 reached 6 mg/L in soluble form, and the purified final product was >96% pure. In Western blot assays, the recombinant IL-6/IL-2 was recognized by polyclonal antibodies against IL-6 and IL-2 of giant panda. The results demonstrated that the protein mixture contained correctly folded IL-2 and IL-6 proteins. A 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide assay demonstrated that IL-6/IL-2 can promote lymphocyte proliferation and differentiation. These data suggest that the fusion protein could be used to develop a novel immunoadjuvant to enhance the immunity of animals against infectious diseases.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Ursidae
/
Interleucina-6
/
Interleucina-2
/
Escherichia coli
Limite:
Animals
Idioma:
En
Revista:
Genet Mol Res
Assunto da revista:
BIOLOGIA MOLECULAR
/
GENETICA
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
China