High level expression of a frog alpha-amidating enzyme, AE-II, in cultured cells and silkworm larvae using a Bombyx mori nuclear polyhedrosis virus expression vector.
A Xenopus laevis peptidyl C-terminal alpha-amidating enzyme (AE-II) gene, modified by deletion of a region encoding the putative membrane-spanning domain and the putative C-terminal cytosolic tail, was expressed in BoMo-15 AIIc insect cells and silkworm larvae using a Bombyx mori baculovirus expression vector system. The expressed enzyme was identified predominantly in the culture medium and the hemolymph of silkworm larvae, indicating successful secretion of the expressed AE-II. The level of recombinant enzyme in the larval hemolymph at 4 days post-infection (40 micrograms/ml) was more than 100-fold the peak levels found in the culture medium (250 ng/ml). The enzyme activity in the larval hemolymph at 4 days post-infection was 3700 units/ml.
Kobayashi
J
J
Department of Insect Genetics and Breeding, National Institute of Sericultural and Entomological Science, Ibaraki, Japan.
Imanishi
S
S
Inoue
H
H
Ohsuye
K
K
Yamaichi
K
K
Tsuruoka
N
N
Tanaka
S
S
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D012831:Bombyx; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004926:Escherichia coli; D005822:Genetic Vectors; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D014982:Xenopus laevis
D004274:DNA, Recombinant; D009097:Multienzyme Complexes; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase