Cloning and Expression of Rabies Virus Glycoprotein Gene into Eukaryotic System

Abstract

Background and Aims: The aim of this study was cloning and expression of rabies virus glycoprotein by a eukaryotic expression plasmid pcDNA3.1(+) in BSR cell line. This construct might be used for a potential DNA vaccine. Materials and Methods: Glycoprotein gene was synthesized and cloned into pBluescript vector and then sub cloned into eukaryotic expression vector (pcDNA3.1(+)). After verification of the cloning, the recombinant plasmid was transfected into BSR cell line (a clone of BHK-21 cell), and its expression was detected by RT-PCR. Results: The authenticity of the recombinant plasmid pcDNA3.1(+)-Gp has been confirmed by a quick check method and restriction endonuclease digestion analysis, and after transfection into eukaryotic cells, the presence of mRNA transcripts was verified by reverse transcriptase-polymerase chain reaction (RT-PCR). Conclusion: This study demonstrated that the construction of eukaryotic expression plasmid for rabies virus glycoprotein is possible. Nevertheless, more work is necessary to develop this kind of vaccine for final use.