Expression of goose interleukin-2 gene in Escherichia coli and isolation of its soluble monomer / 生物工程学报
Chinese Journal of Biotechnology
; (12): 183-187, 2008.
Article
en Zh
| WPRIM
| ID: wpr-276143
Biblioteca responsable:
WPRO
ABSTRACT
Recombinant expression plasmid of pET-28a (+)-goIL-2 was constructed by inserting the goose IL-2 gene without the signal peptide sequence into the prokaryotic expression vector pET-28a (+), and transformed into the bacterial competent E. coli BL21 (DE3) cells for expression. After IPTG induction, an expected protein band with molecular weight of 15.0 kD was observed on SDS-PAGE gel, recognized by monoclonal antibody against goose IL-2 in western-blotting assay. In the pET-28a (+) expression system, much of the recombinant goose IL-2 (rgoIL-2) was found in inclusion bodies with a portion of soluble protein. The monomer and multimers of soluble goose interleukin 2 proteins were observed in native electrophoresis. The rgoIL-2 proteins were purified by Ni-NTA column under a native condition. The rgoIL-2 soluble protein monomer was isolated by a quick protein isolation and purification system of AKTA FPLC and identified by native PAGE. Bioactivity analysis showed that the rgoIL-2 monomer stimulated the proliferation of goose lymphocytes in vitro. This will establish a basis for further study about the biological function and clinical application of goose IL-2.
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Solubilidad
/
Proteínas Recombinantes
/
Cuerpos de Inclusión
/
Interleucina-2
/
Escherichia coli
/
Gansos
/
Genética
/
Metabolismo
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2008
Tipo del documento:
Article