Establishment of a Bombyx mori nucleopolyhedrovirus (BmNPV) hyper-sensitive cell line from the silkworm e21 strain.
Baculoviral expression systems, including those of Autographa californica multiple nucleopolyhedrovirus Bombyx mori nucleopolyhedrovirus (BmNPV), are used for recombinant protein production. Four B. mori-derived (BmN4, Bm5, Bmc140, and Bme21) cell lines were infected with recombinant BmNPV viruses expressing firefly luciferase or EGFP as reporters under the control of a viral polyhedrin promoter. Bme21 exhibited significantly higher (100-fold) luciferase activity than BmN4 and Bm5. With the EGFP reporter protein, Bme21 cells showed a marked increase in the ratio of EGFP-positive cells, reaching 90 % on day 4 post-infection, while Bm5 and BmN4 cells had a slow increase in the ratio of their EGFP-positive population. The viral titer in a supernatant of Bme21 cell culture increased faster than those of Bm5 and BmN4 cells. This susceptibility indicates that the Bme21 cell line is useful for large-scale protein expression using BmNPV.
Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
Jae Man
JM
Lee
Naoya
N
Kawakami
Hiroaki
H
Mon
Hitoshi
H
Mitsunobu
Kazuhiro
K
Iiyama
Satoshi
S
Ninaki
Katsumi
K
Maenaka
Enoch Y
EY
Park
Takahiro
T
Kusakabe
D000818:Animals; D012831:Bombyx / Q000235:genetics* / Q000821:virology*; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D061987:Sf9 Cells; D014162:Transfection
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins