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.
Artigo
em Chinês
| 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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Pichia
/
Proteínas Recombinantes
/
Proteínas Fúngicas
/
Química
/
Reação em Cadeia da Polimerase
/
Regiões Promotoras Genéticas
/
Eletroforese em Gel de Poliacrilamida
/
Flammulina
/
Vetores Genéticos
/
Metabolismo
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2014
Tipo de documento:
Artigo
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