Construction of a Multi-Copy Secretory Expression Vector and Hirudin Expression in Pichia Pastoris
10.3969/j.issn.0253-9896.2010.02.010
- VernacularTitle:多拷贝水蛭素基因在毕赤酵母中的分泌型表达
- Author:
Jingcai MA
;
Xiaona CAO
;
Weili XIONG
;
Jinghua LI
;
Aijun ZUO
;
Bei SUN
;
Dongchun LIANG
- Publication Type:Journal Article
- Keywords:
pichia;
HIRUDIN;
genes;
plasmids;
thrombin
- From:
Tianjin Medical Journal
2010;38(2):112-114,前插2
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct suitable vectors for the secretory expression of hirudin in Pichia pastoris.Methods:The α-facor-hirudin gene was amplified from pPIC9-hirudin by PCR and sub-cloned into PA0815.The multi-copy recombinant plasmid pA0815-(α-Hirudin)n was constructed.The recombinant was transformed into P.pastoris strain GS115 for induction expression and then the activity of secreted products was identified.Results:A new multi-copy vector pA0815-(αHirudin)n was successfully constructed and was capable of secreting recombinant hirudin efficiently,which was confirmed respectively by PCR and SDS-PAGE.The products possessed the activity of thrombin inhibitor.Conclusion:This result offers efficient P.pastoris stains for mass production of biological active hirudin.
