Construction Of Recombinant Plasmids Of G Gene Of Rabies Virus ERA Strain And It's Deleted Mutants And Expression Of Glycoprotein On BHK-21 Cells

The rabies virus glycoprotein (GP) can assume at least three different conformational states :the natural state (N), the activated (A) hydrophobic state and the fusion-inactive (I) state. The entry of rabies virus into host cells requires fusion of the viral envelope with a endosomal membrane. Rabies virus-induced membrane fusion is mediated by viral GP. Fusion is triggered at a low PH, which induced conformational changes of the glycoprotein. It's reported that fusion is occured around PH 5.8-6.0 and is not detected above PH 6.3. Therefore, for determining the fusion domain of G and realize the mechanism of fusion process is great significance for controlling rabies.In this study the a pair primer of G0/G10 were designed and amplified full length of G gene and it's deleted mutants of rabies virus by PCR, then cloned into pMD18-T vector. Then recombinant plasmids pMD-G, pMD-G1G2, pMD-G3G4, pMD-G5G6 and pMD-G7G8 were identified by digestion with restriction endonuclease EcoR I and BstX I. The target gene were extracted and subcloned into eukaryotic expression vector pIREsneo. The recombinant plasmids p-G, p-G1G2, p-G3G4, p-G5G6 and p-G7G8 were constructed. BHK-21 cells were...