The Research On Antigen-characteristics Of Oral RV Gene Vaccine

Rabies is an important public health problem in the whole globe. The death rate of human rabies deaths of China ranked the second place in the world. Rabies is the first serious epidemic diseases in China. Along with the increasing number of dogs, rabies is becoming a big threat to peoples' health. In this study, a strain of Recombinant baculovirus can express glycoprotein of RV in mammalian cell was constructed and researched it as antigeng of gene vaccine which can be taken orally.In this study BHK cells were infected by ERA, and G gene of ERA was amplified by RT-PCR, and then cloned into pMD-18T vector. G gene was digested by restriction endonuclease EcoR I and Pst I, purified and subcloned into recombinant baculovirus transfer vector pAcSG2 which has CMVP as a expression plasmid pAc-G. After detecting by sequencing target gene was the same as G gene of ERA. Expression plasmid pAc-G was transfected into Sf9 to homologous recombine by BD BaculoGold Transfection Kit, and to get recombinant baculovirus. G gene can be amplified from recombinant baculovirus by PCR, and glycoprotein can be detected by IFA. Recombinant baculovirus was purified by plaque purification, and virus titer was 4. 75×107pfu/ml. BHK cells was infected by recombinant baculovirus, and CPE couldn't be seen, but expression of G gene could be detected. All of these indicated that recombinant baculovirus get in this study can invade mammalian cell and express, but can't replicate and harm cell. This study laid a foundation to research antigen of oral RV vaccine.