| RRC ID |
11238
|
| 著者 |
Hong SM, Yamashita J, Mitsunobu H, Uchino K, Kobayashi I, Sezutsu H, Tamura T, Nakajima H, Miyagawa Y, Lee JM, Mon H, Miyata Y, Kawaguchi Y, Kusakabe T.
|
| タイトル |
Efficient soluble protein production on transgenic silkworms expressing cytoplasmic chaperones.
|
| ジャーナル |
Appl Microbiol Biotechnol
|
| Abstract |
Baculovirus expression systems (BES) are widely used for recombinant protein production in lepidopteran cells or larvae. However, even in BES, the insolubility of recombinant proteins sometimes makes their expression difficult. In this study, to improve the solubility and yield of foreign proteins, we constructed transgenic silkworms using silkworm heat-shock proteins, Hsp70 and Hsp40, or Hsc70 and Hsp90 co-chaperone Hop. In these transgenic silkworms, the expression levels of the transgenes were under the control of a UAS.hsp mini-promoter driven by a Gal4NFkBp65 activator. When the transgenic silkworm with HSP70 and 40 (TGS-HSP70/40) was infected with BmNPV carrying mC3d and Gal4NFkBp65 under the control of baculovirus polyhedrin or p10 promoters, respectively, the soluble fraction of the His- or His.GST-tagged mC3d increased significantly. Similarly, the transgenic silkworm with HSC70 and HOP (TGS-HOP7) was effective for the expression of a steroid hormone receptor, USP2. In conclusion, the His-tagged baculovirus expression system featuring the chaperone effect TGS-HSP70/40 and TGS-HOP7 silkworms is effective for increasing the yields of soluble and functional foreign gene products.
|
| 巻・号 |
87(6)
|
| ページ |
2147-56
|
| 公開日 |
2010-8-1
|
| DOI |
10.1007/s00253-010-2617-0
|
| PMID |
20496148
|
| MeSH |
Animals
Animals, Genetically Modified / genetics*
Animals, Genetically Modified / metabolism
Animals, Genetically Modified / virology
Baculoviridae / genetics
Baculoviridae / physiology
Bombyx / genetics*
Bombyx / metabolism
Bombyx / virology
Complement C3d / chemistry
Complement C3d / genetics
Complement C3d / metabolism
Cytoplasm / genetics
Cytoplasm / metabolism*
Gene Expression*
Genetic Vectors / genetics
Genetic Vectors / physiology
Heat-Shock Proteins / genetics*
Heat-Shock Proteins / metabolism
Insect Proteins / chemistry
Insect Proteins / genetics*
Insect Proteins / metabolism
Mice
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism*
Solubility
|
| IF |
3.53
|
| 引用数 |
6
|
|
WOS 分野
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| リソース情報 |
| カイコ |
d17 |
