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

Asit K. Pattnaik, L. Andrew Ball, Alison W. LeGrone, Gail Williams Wertz

Research output: Contribution to journalArticle

222 Scopus citations

Abstract

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 polymerise, 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.

Original languageEnglish (US)
Pages (from-to)1011-1020
Number of pages10
JournalCell
Volume69
Issue number6
DOIs
StatePublished - Jun 12 1992

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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