Expression of Recombinant Nematode Anticoagulant Peptide in Pichia Pastoris / 中国生物工程杂志
China Biotechnology
; (12)2006.
Article
en Zh
| WPRIM
| ID: wpr-685050
Biblioteca responsable:
WPRO
ABSTRACT
Objective: To acquire recombinant nematode anticoagulant peptide ( NAP) with high anticoagulant activity. Methods: Pichia pastoris GS115 strain was transformed with recombinant yeast expression vector pPICS. 5K-rNAP. Expression of rNAP was induced with methanol after the identification of positive strains. NAP expressed in the collected yeast culture supernatant was confirmed with SDS-PAGE and Western blot. The biological activity of the products was validated with PT (prothrombin time) , INR (international normalized ratio) and APTT (activated partial thromboplastin time) , respectively. Results: The yeast strains expressing NAP were identified. The rNAP was secreted into culture supernatant with a molecular weight of about 10 kDa due to glycosylation, which is a little bigger than that predicted (8.7kDa). The anticoagulant efficiency of rNAP was confirmed with the in vitro assays. Conclusion: The recombinant nematode anticoagulant peptide with high biological activity was successfully expressed in Pichia pastoris and can be used in the future development of novel anticoagulant agent.
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Banco de datos:
WPRIM
Idioma:
Zh
Año:
2006
Tipo del documento:
Article