Expression of biologically active recombinant equine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae.
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.
Wu
Donglai
D
National Institute of Animal Health, Tsukuba, Ibaraki 305-085t, Japan.
Murakami
Kenji
K
Liu
Nihong
N
Inoshima
Yasuo
Y
Yokoyama
Takashi
T
Kokuho
Takehiro
T
Inumaru
Shigeki
S
Matsumura
Tomio
T
Kondo
Takashi
T
Nakano
Katsushige
K
Sentsui
Hiroshi
H
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
D000998:Antiviral Agents; D018076:DNA, Complementary; D011994:Recombinant Proteins; D007371:Interferon-gamma