Protection against Autoimmune Diabetes by Silkworm-Produced GFP-Tagged CTB-Insulin Fusion Protein
Figure 2
Identification of the expressed fusion proteins. (a) Fusion protein expression in BmN cells and silkworm B. mori fifth-instar larvae. Left to right: BmNPV CTB-GFP or BmNPV CTB-Ins-GFP 5 days after infection under normal light, blue light, and overlapped. Magnification, 200x. Lower: B. mori fifth-instar normal larvae and larvae infected with recombinant viruses 5 days after infection under ultraviolet radiation. (b) Western blot analysis of fusion protein expression. Lane N: control, lane 1: CTB-Ins-GFP protein, lane 2: CTB-GFP protein. GFP antibody was used as the primary antibody. Silkworm: lane M: molecular weight standard, lane N: normal sample, lane 1: CTB-Ins-GFP protein, lane 2: CTB-GFP protein. Anti-CTB serum was used as the primary antibody. The arrows denote the pentamers (~240, 195 kDa) and monomers (~48, 39 kDa) of the CTB-Ins-GFP and CTB-GFP proteins. (c) ELISA semiquantification analysis of protein expression. Concentrations on days after infection were calculated according to the standard curve of bacterial CTB. Data are presented as the mean concentration ± SD on each day. The experiment was repeated twice.
