Expression of human Versican G1 domain in insect cells

Objective: To produce the recombinant human Versican G1 domain （VG1） in insect cells. Methods: The gene of VG1 which encodes 343 amino acids was inserted into pFastBac1 and the recombinant plasmid was transformed into DH10Bac competent cells for transposition. The recombinant baculovirus were transfected into Sf9 insect cells. The insect cells infected by recombinant baculovirus were determined by Western blot assays. The recombinant protein was purified with Ni-NTA agarose bead column, and its bioactivity was determined with hyaluronan binding assay. Results: The recombinant VG1 was successfully expressed in insect cells. VG1 reacted with the carboxyl-terminal His-tag sequence antibody and with the specific antibody to versican on Western blotting. VG1 bound directly to biotinylated hyaluronan and competitively with unlabeled hyaluronan, indicating that they are folded in a native conformation. Conclusion: Insect cells are suitable to express recombinant human VG1 in high efficiency.