ABSTRACT
To construct a rabies virus mutant, the psi region was replaced by the coding region of human cytochrome c gene, and the coding region for cytoplasmic domain of glycoprotein G was deleted in the full-length of genomic cDNA of rabies virus strain SRV9. The mutant plasmid and the plasmids with N, P, L and G structural proteins of wild type SRV9 were co-transfected into BHK-21 cells. It was shown by IFA that there were many specific fluorescence in the BHK-21 cells, and typical rabies virus virions were observed by electronic microscope. These results demonstrated that the mutant rabies virus was successfully rescued. The genetically modified SRV9 stain has promise to provide invaluable experimental tool to develop attenuated live rabies vaccine.
Subject(s)
Genome, Viral/genetics , Rabies virus/genetics , Animals , Cell Line , Cricetinae , DNA, Complementary/genetics , DNA, Viral/genetics , Humans , Microscopy, Immunoelectron , Mutation , Rabies virus/ultrastructureABSTRACT
A recombinant goatpox virus was constructed in which an enhanced green fluorescent protein gene was inserted under the control of the 11K late promoter, a guanine phosphoribosyltransferase gene was inserted under the control of the 7.5K early/late promoter, and exogenous genes were inserted into an intergenic region between loci gp_24 and gp_24.5 of the recombinant genome. Analysis of protein expression showed that LT cells infected with only the recombinant virus produced specific fluorescence. A comparative growth assay demonstrated the stability of the recombinant virus at the replication level. These results suggest that the intergenic region is not essential for replication of goatpox virus.
