Development of stable cell line expressing secretory form of rabies glycoprotein using a bicistronic messenger

The rabies glycoprotein was expressed as secretory protein in transfected mammalian cells using a bicistronic messenger. The transmembrane domain of rabies glycoprotein was deleted and cloned into a bicistronic expression vector. Upon transfection in mammalian cells, the modified glycoprotein gene was transcribed as bicistronic messenger along with puromycin resistance gene using CMV promoter. The glycoprotein was translated by usual cap dependent translation while puromycin resistance gene was translated by cap-independent manner utilizing IRES from encephalomyocarditis virus. Expressing cells were selected in the presence of antibiotic puromycin and clones expressing glycoprotein were propagated and characterized.