title: [Inducible and constitutive expression of fip-fve from Flammulina velutipes in Pichia pastoris GS115].
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.
authors.forename: Jingwei
authors.initials: J
authors.lastname: Lin
authors.forename: Jia
authors.initials: J
authors.lastname: Jia
authors.forename: Ming
authors.initials: M
authors.lastname: Zhong
authors.forename: Lijing
authors.initials: L
authors.lastname: Chen
authors.forename: Haoge
authors.initials: H
authors.lastname: Li
authors.forename: Zhifu
authors.initials: Z
authors.lastname: Guo
authors.forename: Mingfang
authors.initials: M
authors.lastname: Qi
authors.forename: Lixia
authors.initials: L
authors.lastname: Liu
authors.forename: Tianlai
authors.initials: T
authors.lastname: Li
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D055393:Flammulina / Q000737:chemistry*; D005656:Fungal Proteins / Q000096:biosynthesis*; D005822:Genetic Vectors; D010843:Pichia / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis
chemical_list: C547132:FIP-fve protein, Flammulina velutipes; D005656:Fungal Proteins; D011994:Recombinant Proteins