Efficient soluble protein production on transgenic silkworms expressing cytoplasmic chaperones.
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.
Hong
Sun Mee
SM
Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Fukuoka, Japan.
Yamashita
Jun
J
Mitsunobu
Hitoshi
H
Uchino
Keiro
K
Kobayashi
Isao
I
Sezutsu
Hideki
H
Tamura
Toshiki
T
Nakajima
Hideki
H
Miyagawa
Yoshitaka
Y
Lee
Jae Man
JM
Mon
Hiroaki
H
Miyata
Yoshihiko
Y
Kawaguchi
Yutaka
Y
Kusakabe
Takahiro
T
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D016367:Baculoviridae / Q000235:genetics / Q000502:physiology; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D015933:Complement C3d / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012995:Solubility
D006360:Heat-Shock Proteins; D019476:Insect Proteins; D011994:Recombinant Proteins; D015933:Complement C3d