Infectious defective interfering particles of VSV from transcripts of a cDNA clone.

Pattnaik AK, Ball LA, LeGrone AW, Wertz GW
Cell. 1992 69 (6): 1011-20

PMID: 1318785 · DOI:10.1016/0092-8674(92)90619-n

The generation of infectious defective interfering (DI) particles of vesicular stomatitis virus (VSV) entirely from cDNA clones is reported. Bacteriophage T7 RNA polymerase was used to direct the transcription of a complete negative-stranded genomic RNA from a cDNA clone of a VSV DI RNA in cells simultaneously expressing the five VSV proteins from separately transfected cDNA clones. The negative-stranded transcript was encapsidated with N protein, replicated by the VSV polymerase, and the replicated RNAs were assembled and budded to yield infectious DI virions. No helper VSV was required. Replication occurred at high levels and was assayed by direct biochemical means. An exact 3' terminus of the initial transcript, which was generated by autolytic cleavage using a ribozyme from hepatitis delta virus, was critical for replication.

MeSH Terms (14)

Base Sequence Blotting, Northern Cells, Cultured Defective Viruses DNA In Vitro Techniques Molecular Sequence Data Oligodeoxyribonucleotides RNA, Viral RNA Replicase Transfection Vesicular stomatitis Indiana virus Viral Interference Virus Replication

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