Expression of Recombinant Nematode Anticoagulant Peptide in Pichia Pastoris / 中国生物工程杂志
China Biotechnology
;
(12)2006.
Article
in Chinese
| WPRIM
| ID: wpr-685050
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
China Biotechnology
Year:
2006
Type:
Article
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