[Construction of eukaryotic expression vector of general type in reverse genetic systems of non-segmented negative...

To construct eukaryotic expression vector of general type in reverse genetics systems of non-segmented negative strand RNA viruses, the multiple cloning site of the plasmid pVAX1 was replaced with HamRz cDNA sequence, a 9 sites linker and HdvRz cDNA sequence through the sequential addition of three adapters, the insertion of which could generate the correct 3' and 5' terminal sequences of the primary viral genomic RNA transcript and facilitate the assembly of the complete viral cDNA sequence. The sequences of the three adaptors were correct after identifying by restriction endonuclease digestions and sequencing. The constructed eukaryotic expression vector could not only be used to assemble viral genome, but also provide the basis for establishing the reverse genetic system rapidly.