Production of biologically active recombinant bovine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae.
The full-length bovine interferon-gamma (bIFN-gamma) cDNA, including the secretion signal peptide coding region was recloned into baculovirus transfer vectors pAcYM1 and pBm050. These vectors were co-transfected with Autographa californica nuclear polyhedrosis virus (AcNPV) or Bombyx mori nuclear polyhedrosis virus (BmNPV) DNA into Spodoptera frugiperda cells (SF21AE) and Bombyx mori cells (BmN), respectively. The recombinant viruses, named AcBIFN-gamma and BmBIFN-gamma, were then recovered. Recombinant bIFN-gamma (rbIFN-gamma) was accumulated in the culture fluid of AcBIFN-gamma-infected Trichoplusia ni cells and BmBIFN-gamma-infected silkworm larvae. These rbIFN-gamma forms were shown to be glycosylated 20 and 22 kDa proteins as confirmed by SDS-PAGE and tunicamycin treatment. These products were sensitive to cystein proteinase. Both rbIFN-gamma proteins, showed high-level biological activities by plaque reduction assay using vesicular stomatitis virus, and MHC class II antigen induction on bovine macrophage cells.
Murakami
K
K
National Institute of Animal Health, 3-1-1 Kannondai, Tsukuba, Ibaraki, 305-0856, Japan.
Uchiyama
A
A
Kokuho
T
T
Mori
Y
Y
Sentsui
H
H
Yada
T
T
Tanigawa
M
M
Kuwano
A
A
Nagaya
H
H
Ishiyama
S
S
Kaki
H
H
Yokomizo
Y
Y
Inumaru
S
S
D000818:Animals; D001483:Base Sequence; D012831:Bombyx; D002417:Cattle; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006031:Glycosylation; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D007814:Larva; D008970:Molecular Weight; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011480:Protease Inhibitors / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D018411:Spodoptera
D018076:DNA, Complementary; D004274:DNA, Recombinant; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D007371:Interferon-gamma