Construction of an expression vector for improved secretion of canine IL-18.
To construct a vector for caspase-1 independent expression of canine IL-18, the signal sequence of canine IL-12p40 was fused to the sequence of mature IL-18 on the NdeI restriction site which is located at the 3' end of the signal sequence. The resulting vector expressed coding protein from transfected mammalian cells. The expressed protein was shown to have IL-18 bioactivity in a INF-gamma-inducing assay. These results suggest that the expression vector is the desired tool for advancement of dendritic cell (DC)-based cancer therapy, provided that the vector can successfully be transfected into dendritic cells. We propose a simple and widely applicable method for providing the signal sequence.
Sugiura
Kikuya
K
Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho Naka-ku, Sakai City, Osaka 599-8531, Japan. sugiura@vet.osakafu-u.ac.jp
Akazawa
Takashi
T
Fujimoto
Mariko
M
Wijewardana
Viskam
V
Mito
Kai
K
Hatoya
Shingo
S
Taketani
Shigeru
S
Komori
Masayuki
M
Inoue
Norimitsu
N
Inaba
Toshio
T
D000595:Amino Acid Sequence; D000704:Analysis of Variance; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004285:Dogs; D005822:Genetic Vectors / Q000235:genetics*; D053762:Interleukin-12 Subunit p40 / Q000235:genetics; D020382:Interleukin-18 / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000662:veterinary
D017931:DNA Primers; D053762:Interleukin-12 Subunit p40; D020382:Interleukin-18