Prokaryotic expression, purification and identification of recombinant human atrial natriuretic peptide / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1273-1285, 2016.
Artículo
en Chino
| WPRIM
| ID: wpr-310540
ABSTRACT
In order to improve the expression of recombinant human atrial natriuretic peptide (ANP), a new plasmid (pET28a(+)/ANP₃) containing 3 tandem ANP genes with lysine codon as the interval linker, was constructed. Target gene was transformed into Escherichia coli BL21 (DE3) and induced by IPTG, about 60% of the total-cell-protein was the target protein, His₆-ANP₃. After denaturation and refolding, it was digested by Endoproteinase Lys-C and Carboxypeptidase B (CPB) and then purified by a series of purification processes, about 16 mg purified ANP monomer could be obtained from one liter bacteria broth of shaking culture. Ultimately, the purity of protein was above 90% determined by UPLC and Tricine SDS-PAGE, its molecular weight was 3 080 Da according to LC-MS identification and it was proved to be equivalent to the reference product by ELISA. The use of tandem gene expression can provide a new possible model for the expression of other peptide drugs.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Péptidos
/
Plásmidos
/
Proteínas Recombinantes de Fusión
/
Metaloendopeptidasas
/
Expresión Génica
/
Factor Natriurético Atrial
/
Electroforesis en Gel de Poliacrilamida
/
Escherichia coli
/
Genética
/
Metabolismo
Tipo de estudio:
Estudio pronóstico
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2016
Tipo del documento:
Artículo
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