Infectious measles virus from cloned cDNA

I. Ballart, D. Eschle, R. Cattaneo, A. Schmid, M. Metzler, J. Chan, S. Pifko-Hirst, S. A. Udem, M. A. Billeter

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

The study of measles virus (MV) and of negative strand RNA viruses in general has been hampered by the lack of an experimental system for genetic manipulation. Here we describe a procedure for generating infectious MV from cloned MV cDNA. First we assembled a genetically marked DNA copy of the MV genome in plasmids, under the control of phage T3 or T7 promoters, allowing production of transcripts almost identical to the MV genome or antigenome. Incubation of these linearized plasmid DNAs with the appropriate phage polymerase and only two ribonucleoside triphosphates yielded committed transcription complexes. Microinjection of these complexes into the cytoplasm of helper cells which provide the proteins necessary for MV genome encapsidation and transcription/replication, reproducibly give rise to lytic MVs. The transcripts of one of these viruses were analysed by sequencing after reverse transcription followed by DNA amplification, and found to contain the genetic tags, The described procedure permits the analysis of a negative strand RNA virus with the same genetic tools previously applicable only to positive strand RNA viruses and retroviruses.

Original languageEnglish (US)
Pages (from-to)379-348
Number of pages32
JournalEMBO Journal
Volume9
Issue number2
StatePublished - Feb 20 1990

Keywords

  • Genome reconstruction
  • RNA virus
  • infectious cDNA
  • measles virus
  • negative strand RNA

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Fingerprint Dive into the research topics of 'Infectious measles virus from cloned cDNA'. Together they form a unique fingerprint.

  • Cite this

    Ballart, I., Eschle, D., Cattaneo, R., Schmid, A., Metzler, M., Chan, J., Pifko-Hirst, S., Udem, S. A., & Billeter, M. A. (1990). Infectious measles virus from cloned cDNA. EMBO Journal, 9(2), 379-348.