Construction and Immune Effect of Recombinant Retrovirus Containing Glycoprotein Gene of Rabies Virus

Objective To construct a recombinant retrovirus containing the glycoprotein(GP)gene of rabies virus and determine its immune effect in mice. Methods Plasmid pVAX-G containing the GP gene of rabies virus SRV9 strain was digested with restriction endonuclease, and the obtained gene fragment was cloned into retrovirus vector. The constructed recombinant plasmid pLNCX-G was transfected to packaging cell line PA317 in mediation of liposome, and positive cell clones were screened for cloning and amplification. BHK cells were infected with virus particles, in which the integration and transcription of GP gene were determined by PCR and RT-PCR. The titer of retrovirus was determined with NIH3T3 cells. KM mice were immunized with the constructed recombinant retrovirus, and determined for serum neutralizing antibody titer against rabies virus by fluorescent antibody virus neutralization(FAVN)test. Results Restriction analysis proved that recombinant plasmid pLNCX-G was constructed correctly. The GP gene of rabies virus was integrated to the genome of BHK cells and transcribed normally. The virus titer reached 1. 2 × 104 CFU / ml at the most. The positive rates of neutralizing antibody against rabies virus induced in mice by intramuscular and intraperitoneal injections were 6. 7% and 86. 7%, while the effective protection rates were 0 and 76. 9% respectively. Conclusion The recombinant retrovirus containing the GP gene of rabies virus SRV9 strain was successfully constructed, and a certain neutralizing antibody titers were induced in mice, which laid a foundation of further study.