Inducible and constitutive expression of fip-fve from Flammulina velutipes in Pichia pastoris GS115 / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 464-471, 2014.
Article
in Chinese
| WPRIM
| ID: wpr-279503
ABSTRACT
We transformed the fip-fve gene into Pichia pastoris GS115 for inducible and constitutive expression to obtain feasible bioactvie recombinant Fip-fve. The fip-fve gene was cloned from Flammulina velutipes fruting body by PCR and ligated to pPIC9 to construct inducible expression vector pPIC9-FIP-fve, and promotor pgap was used to replace the paox1 to construct constitutive expression vector pPIC9-PGAP-FIP-fve. These two vectors were used to transform P. pastoris by PEG method. The fip-fve was expressed after histamine-absence screening and yeast colony PCR. The inducible expression level reached 158.2 mg/L at the fourth day and the constitutive expression level was 46.3 mg/L and 29.5 mg/L using glucose and glycerol, respectively. The SDS-PAGE and Western blotting both proved the correctness of rFip-fve, and the hemagglutination test indicats the rFip-fve's bioactivity.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pichia
/
Recombinant Proteins
/
Fungal Proteins
/
Chemistry
/
Polymerase Chain Reaction
/
Promoter Regions, Genetic
/
Electrophoresis, Polyacrylamide Gel
/
Flammulina
/
Genetic Vectors
/
Metabolism
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2014
Type:
Article
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