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. Author manuscript; available in PMC: 2011 Jun 9.
Published in final edited form as: Biotechniques. 2010 Jul;49(1):513–518. doi: 10.2144/000113450

Figure 1. Construction and Expression of G-C-T Fusion Protein.

Figure 1

(a) Design of the recombinant fusion protein. The G-C-T fusion protein is linked via a dithiocyclopeptide linker based on the structure of somatostatin modified to contain a thrombin-specific sequence, PRS. Two cysteinyl-residues on somatostatin naturally form a disulfide bond. (b) Plasmid encoding the G-C-T fusion protein was constructed based on the mammalian expression vector pcDNA3.0. (c) Recombinant G-C-T was analyzed by western blot using anti-human G-CSF or serum Tf primary antibody, with horseradish peroxidase-conjugated anti-rabbit or anti-goat, respectively, IgG secondary antibody. The peroxidase activity was detected by enhanced chemiluminescence using the ECL substrate system.