title: Expression of biologically active recombinant equine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae.
abstract: The full-length equine interferon-gamma (eIFN-gamma) cDNA, including the secretion signal peptide coding region, was recloned into baculovirus transfer vector pAcYM1. This vector was co-transfected with Autographa californica nuclear polyhedrosis virus DNA or hybrid nuclear polyhedrosis virus DNA into Spodoptera frugiperda cells. The recombinant viruses, named AcEIFN-gamma and HyEIFN-gamma, were then recovered. Recombinant eIFN-gamma (reIFN-gamma) was accumulated in the culture fluid of the AcEIFN-gamma or HyEIFN-gamma infected Tricoplusia ni -derived cell line, BTI TN 5B1-4, and hemolymph of HyEIFN-gamma infected silkworm larvae. These reIFN-gamma forms were shown to be 14, 16, 18 and 20kDa proteins, and glycosylated as confirmed by SDS-PAGE and tunicamycin treatment. Both reIFN-gamma proteins, showed high-level biological activities to vesicular stomatitis virus by cytopathic effect reduction assay, and MHC class II antigen induction on the equine fetal kidney-78 cell line.
authors.affiliation: National Institute of Animal Health, Tsukuba, Ibaraki 305-085t, Japan.
authors.forename: Donglai
authors.initials: D
authors.lastname: Wu
authors.forename: Kenji
authors.initials: K
authors.lastname: Murakami
authors.forename: Nihong
authors.initials: N
authors.lastname: Liu
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Inoshima
authors.forename: Takashi
authors.initials: T
authors.lastname: Yokoyama
authors.forename: Takehiro
authors.initials: T
authors.lastname: Kokuho
authors.forename: Shigeki
authors.initials: S
authors.lastname: Inumaru
authors.forename: Tomio
authors.initials: T
authors.lastname: Matsumura
authors.forename: Takashi
authors.initials: T
authors.lastname: Kondo
authors.forename: Katsushige
authors.initials: K
authors.lastname: Nakano
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sentsui
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D012831:Bombyx; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006736:Horses; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D007814:Larva; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011994:Recombinant Proteins; D018411:Spodoptera; D014721:Vesicular stomatitis Indiana virus / Q000472:pathogenicity
chemical_list: D000998:Antiviral Agents; D018076:DNA, Complementary; D011994:Recombinant Proteins; D007371:Interferon-gamma