A plasmid-based reverse genetics system for influenza A virus.
作者： Stephan Pleschka , R Jaskunas , Othmar G Engelhardt , T Zürcher , Peter Palese
DOI: 10.1128/JVI.70.6.4188-4192.1996
关键词:
摘要: A reverse genetics system for negative-strand RNA viruses was first successfully developed influenza viruses. This technology involved the transfection of in vitro-reconstituted ribonucleoprotein (RNP) complexes into virus-infected cells. We have now a method that allows intracellular reconstitution RNP from plasmid-based expression vectors. Expression viral RNA-like transcript is achieved plasmid containing truncated human polymerase I (polI) promoter and ribozyme sequence generates desired 3' end by autocatalytic cleavage. The polI-driven cotransfected 293 cells with polII-responsive plasmids express PB1, PB2, PA, NP proteins. exclusively plasmid-driven results efficient transcription replication reporter study cis- trans-acting signals virus RNAs. Using this system, we also been able to rescue synthetic neuraminidase gene recombinant virus. represents convenient alternative previously established system.
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