Expression of platelet collagen receptor-glycoprotein VI fragment in E. coli and its biological activities / 中国实验血液学杂志
Journal of Experimental Hematology
;
(6): 304-308, 2005.
Artículo
en Chino
| WPRIM
| ID: wpr-356572
ABSTRACT
This study was aimed to further investigate the function of platelet collagen receptor-glycoprotein VI and to screen its specific inhibitor. The extracellular domain of platelet glycoprotein VI (GPVI) in E. coli was expressed by recombinant technology, the extracellular domain cDNA of GPVI was amplified from pBluescript KS(-)-GPVI plasmid by PCR. Proved by sequencing, the expression vector pET-20b(+)-GPVI was constructed, which was then transformed into E. coli (BL21(DE3)pLysS) and induced by IPTG. The recombinant GPVI was purified on Ni-NTA resin column and renatured in PBS containing GSH and GSSG. The anti-penta His McAb and anti-GPVI polyclonal antibody were used to identify the recombinant GPVI in Western blotting. Collagen binding test was conducted to investigate the biological activity of recombinant GPVI. The results showed that the recombinant GPVI was expressed in E. coli and successfully purified, which was confirmed to be similar to the native GPVI in Western blotting. The recombinant GPVI can bind the type I collagen in dose-dependent manner. In conclusion, the recombinant GPVI can be achieved in E. coli and restore its native characteristics after renaturation.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Unión Proteica
/
Plaquetas
/
Proteínas Recombinantes
/
Glicoproteínas de Membrana Plaquetaria
/
Western Blotting
/
Integrina alfa2beta1
/
Receptores de Colágeno
/
Escherichia coli
/
Genética
/
Metabolismo
Tipo de estudio:
Estudio pronóstico
Límite:
Humanos
Idioma:
Chino
Revista:
Journal of Experimental Hematology
Año:
2005
Tipo del documento:
Artículo
Similares
MEDLINE
...
LILACS
LIS