Expression of equine interleukin-18 by baculovirus expression system and its biologic activity.
The equine interleukin-18 (IL-18) cDNA that contains the coding sequence was cloned and a recombinant baculovirus, named AcEIL-18, was constructed. The recombinant protein of the equine IL-18 was expressed by AcEIL-18 and its expression was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Insect cells infected with AcEIL-18 secreted a precursor IL-18 with 24 kilo dalton (kDa) into the culture supernatant. Western blot analysis showed that mature equine IL-18 about 18 kDa was also confirmed without co-expression of caspase-1. Culture supernatant from AcEIL-18 infected cells showed a synergistic effect with recombinant human interleukin-12 for induction of interferon-gamma gene expression in equine peripheral mononuclear cells, indicating that the recombinant equine IL-18 expressed in this study also has biological activity without any treatment.
Wu
Donglai
D
National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
Murakami
Kenji
K
Liu
Nihong
N
Konishi
Misako
M
Muneta
Yoshihiro
Y
Inumaru
Shigeki
S
Kokuho
Takehiro
T
Sentsui
Hiroshi
H
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D003470:Culture Media; D015870:Gene Expression; D006736:Horses / Q000097:blood / Q000276:immunology*; D007371:Interferon-gamma / Q000096:biosynthesis; D020382:Interleukin-18 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008970:Molecular Weight; D015202:Protein Engineering; D011498:Protein Precursors / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000276:immunology
D003470:Culture Media; D020382:Interleukin-18; D011498:Protein Precursors; D011994:Recombinant Proteins; D007371:Interferon-gamma