title: Molecular cloning and expression of a fungal immunomodulatory protein, FIP-fve, from Flammulina velutipes.
abstract: FIP-fve, a fungal immunomodulatory protein, was isolated from the fruiting bodies of the edible mushroom, Flammulina velutipes. FIP-fve was shown to stimulate blast-forming activity of human peripheral blood lymphocytes and gene expression of interleukin-2, interferon-gamma and tumor necrosis factor-alpha. Repeated administration of FIP-fve to mice inhibits the Arthur and systemic anaphylaxis reactions. FIP-fve cDNA was cloned and sequenced, and the amino acid sequence of FIP-fve deduced from the nucleotide sequence is identical to that previously determined by protein sequencing. FIP-fve cDNA was amplified by polymerase chain reaction, ligated into the expression vector, pGEX-2T, and expressed in Escherichia coli as a fusion protein of glutathione S-transferase (GST) and FIP-fve. The GST-FIP-fve fusion protein was soluble, and the yield of recombinant FIP-fve was about 5 mg/L of induced culture. The recombinant FIP-fve was obtained by cleaving the GST-FIP-fve fusion protein with thrombin and purifing to homogeneity. The recombinant FIP-fve had about 50% of the immunomodulatory activity of the native FIP-fve.
authors.affiliation: Institute of Biochemistry, College of Medicine, National Taiwan University, Taipei, ROC.
authors.forename: J L
authors.initials: JL
authors.lastname: Ko
authors.forename: S J
authors.initials: SJ
authors.lastname: Lin
authors.forename: C I
authors.initials: CI
authors.lastname: Hsu
authors.forename: C L
authors.initials: CL
authors.lastname: Kao
authors.forename: J Y
authors.initials: JY
authors.lastname: Lin
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D018076:DNA, Complementary / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D037102:Lectins; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins
chemical_list: D003062:Codon; D018076:DNA, Complementary; D005656:Fungal Proteins; D037102:Lectins; D011994:Recombinant Proteins