[Expression of glycoprotein gene of the rabies virus 3aG strain by E-3 deleted adenovirus recombinant].

OBJECTIVE: To analyze Chinese rabies virus vaccine strain 3aG glycoprotein (GP) gene and further produce GP by E3-deleted human adenovirus recombinant. METHODS: Chinese rabies virus vaccine strain 3aG glycoprotein gene was cloned by RT-PCR and its sequence was determined by DNA sequencing. Cotransfection was performed to obtain adenovirus recombinant. The expressed glycoprotein was examined by ELISA and its immunogenicity was evaluated by testing neutralizing antibody level of mice inoculated with the recombinant virus. RESULTS: DNA sequencing showed that the open reading frame of GP gene contains 1,575 nucleotides and five of the deduced amino acids are different from the previous report. The recombinant adenovirus containing GP gene in E3 region was obtained by cotransfecting 293 cells and rounds of plaque purification. ELISA assay demonstrated that the GP gene can be efficiently expressed and rapid fluorescent focus inhibition test (RFFTT) showed it can also elicit GP specific neutralizing antibody. CONCLUSIONS: Chinese rabies virus vaccine strain 3aG glycoprotein was successfully expressed by E3-deleted human adenovirus recombinant and specific neutralizing antibody can be elicited in mice after immunization by the recombinant.