Secretory expression of BPI_(m23) recombinant bactericidal protein in Pichia pastoris / 中国免疫学杂志
Chinese Journal of Immunology
; (12)1985.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-675583
Responsible library:
WPRO
ABSTRACT
Objective:
To express and secrete rBPI m23 bactericidal protein in Pichia pastoris expression system.Methods:
pPICZ? synBPI m600 recombinant expression vector was constructed with prepared vectors (pUC18 synBPI 414 and PBV220 BPI m420 ) according to Molecular Cloning Laboratory Manual. The Pichia pastoris GS115 were electroporated with linearized pPICZ? synBPI m600 vectors , positive clones were screened on low salt LB medium with Zeocin , then the positive clones were identified by PCR and Mut phenotype analysis. rBPI m23 was induced to express in BMMY ,purified by SP Sepharose beads , identified by SDS PAGE and Western blot and measured with BCA method.Results:
①pPICZ? synBPI m600 expression vector was successfully constructed.②Pichia pastoris GS115 strains integrated with synBPI m600 stably were obtained.③SDS PAGE showed the molecular weight of the recombinant protein was approximate 23 kD,and Western blot confirmed that the protein could bind to the anti BPI antibody specially.④The supernate protein concentration was about 2 9 mg/L.Conclusion:
rBPI m23 was expressed and secreted effectively in Pichia pastoris GS115.
Full text:
Available
Database:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Immunology
Year:
1985
Document type:
Article