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Chinese Journal of Biotechnology ; (12): 464-471, 2014.
Article de Chinois | WPRIM | ID: wpr-279503

RÉSUMÉ

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.


Sujet(s)
Électrophorèse sur gel de polyacrylamide , Flammulina , Chimie , Protéines fongiques , Vecteurs génétiques , Pichia , Métabolisme , Réaction de polymérisation en chaîne , Régions promotrices (génétique) , Protéines recombinantes
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