Optimization of prokaryotic expression condition and purification of anti-cancer protein NOR1 in E.coli / 中南大学学报(医学版)
Journal of Central South University(Medical Sciences)
;
(12): 610-615, 2011.
Artigo
em Chinês
| WPRIM
| ID: wpr-814542
ABSTRACT
OBJECTIVE@#To optimize the induction condition of human NOR1 gene expression in E.coli. and purify NOR1 recombinant proteins.@*METHODS@#A full-length cDNA of human NOR1 was inserted into the corresponding region of pET28b expression vector to yield recombinant prokaryotic expression vector pET28b-NOR1. The prokaryotic expression vector pET28b-NOR1 was introduced into the bacterial host E.coli Rosettablue(DE3). Recombinant NOR1 protein was induced at different conditions. Induction condition was optimized to obtain high yield of recombinant protein. At last, the recombinant NOR1 protein was purified by Ni-IDE chromatography resin.@*RESULTS@#Recombinant NOR1 protein was induced by IPTG in a dose-dependent manner. Increase of kanamycin concentration and induction temperature resulted in high yield of recombinant protein. The most recombinant protein was found in inclusion bodies. The recombinant His-NOR1 protein was purified with Ni-IDE chromatography resin under denature condition.@*CONCLUSION@#IPTG, kanamycin concentration and temperature can affect the expression of recombinant NOR1 protein in pET28b system. High yield of recombinant NOR1 protein is achieved by inducing 1 mmol/L IPTG and 200 μg/mL kanamycin at 37 degree. Recombinant His-NOR1 protein with high purity is purified.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas de Membrana Transportadoras
/
Farmacologia
/
Proteínas Recombinantes
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Proteínas Supressoras de Tumor
/
Escherichia coli
/
Vetores Genéticos
/
Genética
/
Isopropiltiogalactosídeo
Limite:
Humanos
Idioma:
Chinês
Revista:
Journal of Central South University(Medical Sciences)
Ano de publicação:
2011
Tipo de documento:
Artigo
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