RRC ID |
3449
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著者 |
Miao Y, Zhang Y, Nakagaki K, Zhao T, Zhao A, Meng Y, Nakagaki M, Park EY, Maenaka K.
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タイトル |
Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system.
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ジャーナル |
Appl Microbiol Biotechnol
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Abstract |
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.
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巻・号 |
71(2)
|
ページ |
192-9
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公開日 |
2006-6-1
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DOI |
10.1007/s00253-005-0127-2
|
PMID |
16158284
|
MeSH |
Amino Acid Sequence
Animals
Baculoviridae / genetics*
Base Sequence
Biotechnology / methods*
Blotting, Western
Bombyx / metabolism
Bombyx / virology*
Cloning, Molecular
DNA / chemistry
DNA / genetics
Molecular Sequence Data
Polymerase Chain Reaction
Proteins / genetics
Proteins / metabolism*
Recombinant Proteins / biosynthesis
Recombinant Proteins / genetics
Spiders / genetics*
Spiders / metabolism
|
IF |
3.53
|
引用数 |
68
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WOS 分野
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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リソース情報 |
カイコ |
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