Figure 1 | Scientific Reports

Figure 1

From: Co-expression of BirA with biotin bait achieves in vivo biotinylation of overexpressed stable N-glycosylated sRAGE in transgenic silkworms

Figure 1

Expression of sRAGE in MSGs of transgenic silkworms. (A) Upper panel: Schematic diagrams of the domain structure of the extracellular domain of human RAGE. Numbers correspond to the amino acid (aa) sequence of full-length hRAGE, a 404-amino acid protein composed of an extracellular domain (aa 23–332). C represents cysteine residues involved in S-S linkages. Lower panel: Amino acid sequence of sRAGE expressed in this study. Underlined amino acids represent the RAGE region; bold indicates the BioEase tag; and boxes represent the potential N-glycosylation sites. (B) Generation of transgenic silkworms expressing the sRAGE gene. Adults transgenic silkworms harboring the sRAGE gene or GAL4 gene were mated. The MSGs were isolated from F1 larvae, and total proteins were extracted. (C) CBB staining of SDS-PAGE. The protein lysates extracted from MSGs were separated by SDS-PAGE followed by staining with CBB. (D) Detection with anti-His-tag antibodies. Lysates were separated with SDS-PAGE, followed by western blotting with anti-His-tag antibodies. The protein lysates extracted from MSGs of transgenic silkworms harboring only the Ser1-GAL4 construct were used as negative controls (−). (E) Detection with anti-RAGE antibodies. The numbers above the gel or blots indicate the line number of the transgenic strain. Numbers on the left indicate the molecular mass (kDa).

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