Production of recombinant sarcotoxin IA in Bombyx mori cells.
A cDNA for sarcotoxin IA, an antibacterial protein of Sarcophaga peregrina (fleshfly), was inserted into a silkworm baculovirus vector and expressed in Bm-N cells, a line of Bombyx mori cells. When a cysteine proteinase inhibitor, p-chloromercuribenzenesulphonic acid, was present in the culture medium, a significant amount of recombinant sarcotoxin IA accumulated, but without this reagent the product seemed to be degraded in this system. The C-terminus of the recombinant sarcotoxin IA seemed to be glycine, not amidated arginine as found in authentic sarcotoxin IA. Probably, Bm-N cells lack the C-terminal alpha-amidation enzyme.
Yamada
K
K
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Nakajima
Y
Y
Natori
S
S
D000818:Animals; D012831:Bombyx; D002460:Cell Line; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004175:Diptera / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D007301:Insect Hormones / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D019476:Insect Proteins; D008970:Molecular Weight; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014162:Transfection
D007301:Insect Hormones; D019476:Insect Proteins; D011994:Recombinant Proteins; C054733:sarcotoxin IA protein, Sarcophaga peregrina