Expression of secreted form of rabies virus glycoprotein from stably transfected mammalian cell line

Rabies virus glycoprotein was expressed as secretory glycoprotein from transfected Madin Darby bovine kidney (MDBK) cells. The N-terminal viral signal peptide was replaced with CD33 signal peptide for efficient secretion in non-neuronal mammalian cells. The 60 amino acid C-terminal hydrophobic anchor (transmembrane) and cytoplasmic domains were also replaced with polyhistidine tag, which facilitated purification of expressed protein. The stably transfected cells were selected in the presence of G418 and cell line was maintained. From the cell culture supernatant, the recombinant secretory form of glycoprotein was purified using Ni-agarose affinity chromatography utilizing C-terminal polyhistidine tag. The purified recombinant glycoprotein was characterized in SDS-PAGE and western blot.