Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system.
Bombyx mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system (BES) has a lot of advantages such as high expression efficiency, convenience, and low feeding cost. In this report, we used a recently developed BmNPV bacmid, which could infect both B. mori cell lines and silkworm larvae. The results showed it takes only 7 to 10 days to generate recombinant baculovirus and permit the rapid isolation from small-scale cultures and then use it to transfect B. mori cell lines, compared to traditional homologous recombination method, which needs at least 40 days for multiple rounds of purification and amplification of viruses. Using this BES, we expressed a recombinant spider flagelliform protein in BmN cell line, which was around 37 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. The BmNPV bacmid system using silkworm would be very attractive for expression of target proteins.
Miao
Yungen
Y
College of Animal Sciences, Zhejiang University, Hangzhou, China. miaoyg@zju.edu.cn
Zhang
Yuansong
Y
Nakagaki
Koichi
K
Zhao
Tianfu
T
Zhao
Aichun
A
Meng
Yan
Y
Nakagaki
Masao
M
Park
Enoch Y
EY
Maenaka
Katsumi
K
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods*; D015153:Blotting, Western; D012831:Bombyx / Q000378:metabolism / Q000821:virology*; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013112:Spiders / Q000235:genetics* / Q000378:metabolism
C404864:FLAG protein, Nephila; D011506:Proteins; D011994:Recombinant Proteins; D004247:DNA