Wuhan Institute of Biological Ihoducts Co.

Wuhan, China

Wuhan Institute of Biological Ihoducts Co.

Wuhan, China

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Xie T.-B.,Wuhan Institute of Biological Ihoducts Co. | Ming P.-G.,Wuhan Institute of Biological Ihoducts Co. | Tang F.,Wuhan Institute of Biological Ihoducts Co. | Huang S.-J.,Wuhan Institute of Biological Ihoducts Co. | And 4 more authors.
Chinese Journal of Biologicals | Year: 2015

Objective To reconstruct the reverse genetic system of rabies virus CTN strain and rescue the modified recombinant rabies virus. Methods The amino acids at sites 194 and 333 of G protein of CTN strain were mutated by site-directed mutagenesis technology. The mutated G gene was used as a substitute for the former G gene, based on which a novel system carrying a single, double and triple mutated G gene was constructed. The reconstructed full-length cDNA and auxiliary plasmids were co-transfected to BSR cells, and the rescued vims was identified by direct immunofluorescence assay (DFA) and RT-PCR. Results The mutation of G gene (194 Asn → Ser, 333 Gln → Glu) was observed. The reverse genetic systems carrying a single, double and triple mutated G gene were constructed respectively, and the recombinant virus was rescued. Conclusion The reverse genetic system of rabies virus CTN strain was successfully modified, and the recombinant virus was rescued, which laid a foundation of developing a novel, safe, stable and effective rabies vaccine.

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