Expression of glycoprotein gene of the rabies virus 3aG strain by E-3 deleted advenovirus recombinant

Abstract

Objective To analysis Chinese rabies vibies virus vaccine strain 3aG glycoprotein(GP) gene and furtherproduce GP by E3-deleted human adenovirus recombinant.Methods Chinese rabies virus vaccine strain 3aG glyco-protein gene was cloned by RT-PCR and its sequence was determined by DNA sequencing.Cotransfection was per-formed to obtain adenovirus recombinant.The expressed glycoprotein was examined by ELISA and its immunogenicitywas evaluated by testing neutralizing antibody level of mice inoculated with the recombinant virus.Results DNA se-quencing showed that the open reading frame of GP gene contains 1 575 nucleotides and five of the deduced amino acidsare different from the previous report.The recombinant adenovirus containing GP gene in E3 region was obtained by co-transfecting 293 cells and rounds of plaque purification. ELISA assay demonstrated that the GP gene can be efficientlyexpressed and rapid fluorescent focus inhibition test (RFFTT) showed it can also elicit GP specific neuralizing antibody.Conclusion Ciinese rabies vurus vaccine strain 3aG glycoproteein was successfully expresssed by E3 - deleted humanadenovirus recombinant snd sprvigic nrutralizing snyinody can be elicited in mice sfter immunixsyion by the recombinant. Key words: Rabies glycoprotein; Seqyence abalyis; Reconbubabt adebivurys; Neulilizng antibody