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....

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2006-06, Vol.71 (2), p.192-199
Main Authors: Miao, Y, Zhang, Y, Nakagaki, K, Zhao, T, Zhao, A, Meng, Y, Nakagaki, M, Park, E.Y, Maenaka, K
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Araneae
Baculoviridae - genetics
Baculovirus
Base Sequence
Biological and medical sciences
Biotechnology
Biotechnology - methods
Blotting, Western
Bombyx - metabolism
Bombyx - virology
Bombyx mori
Bombyx mori NPV
Cloning, Molecular
DNA - chemistry
DNA - genetics
Enzymes
Fundamental and applied biological sciences. Psychology
Larvae
Molecular Sequence Data
Nuclear polyhedrosis virus
Polymerase Chain Reaction
Proteins
Proteins - genetics
Proteins - metabolism
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Silk
Spiders - genetics
Spiders - metabolism
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Summary: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.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-005-0127-2