Your browser doesn't support javascript.
loading
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.
Artigo em Chinês | 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.
Assuntos

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Peptídeos / Plasmídeos / Proteínas Recombinantes de Fusão / Metaloendopeptidases / Expressão Gênica / Fator Natriurético Atrial / Eletroforese em Gel de Poliacrilamida / Escherichia coli / Genética / Metabolismo Tipo de estudo: Estudo prognóstico Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Biotechnology Ano de publicação: 2016 Tipo de documento: Artigo

Similares

MEDLINE

...
LILACS

LIS

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Peptídeos / Plasmídeos / Proteínas Recombinantes de Fusão / Metaloendopeptidases / Expressão Gênica / Fator Natriurético Atrial / Eletroforese em Gel de Poliacrilamida / Escherichia coli / Genética / Metabolismo Tipo de estudo: Estudo prognóstico Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Biotechnology Ano de publicação: 2016 Tipo de documento: Artigo