Expression of von Willebrand factor-A1 domain in E coli and it's the inhibitory effect on platelet aggregation / 中国病理生理杂志
Chinese Journal of Pathophysiology
; (12)1986.
Article
en Zh
| WPRIM
| ID: wpr-521949
Biblioteca responsable:
WPRO
ABSTRACT
AIM: To further investagate the mechanism of thrombus formation and develop a new remedy of anti-thrombus formation. METHODS: The amplified DNA fragment of vWF-A1 domain was inserted into expression vector with 6?his taq (pQE-31), the recombinant expression vect or was transformed into E coli (strain M15) and induced by IPTG. The recombinant fragment, comprising residues 449-728 of mature vWF subunit, designate rvWF-A1. It was purified by Ni-NTA agarose column and renatured by Tris buffer containin g GSH and GSSG. FACS and platelet aggregometer were employed to analyse the rvWF -A1 function of binding to platelet glycoprotein Ib and inhibiting ristocetin-in duced platelet aggregation. RESULTS: The rvWF-A1 was expressed successfully in E coli, comin g up to 30% of total bacterial protein. Its purify was over 95% through Ni-NTA a garose. It was identified to have ability to bind to GPIb, its biologic activity to inhibit ristocetin-induced platelet aggregation was observed, and the inhibi tive rate was 84 7%. CONCLUSION: The above results indicated that high-level expressi on of rvWF-A1 was successfully achieved in E coli and rvWF-A1 may be an effectiv e antithromotic agent in preventing thrombus formation.
Texto completo:
1
Base de datos:
WPRIM
Tipo de estudio:
Prognostic_studies
Idioma:
Zh
Revista:
Chinese Journal of Pathophysiology
Año:
1986
Tipo del documento:
Article