Infectious measles virus from cloned cDNA

Isidro Ballart, Daniel Eschle, Roberto Cattaneo, Anita Schmid, Martina Metzler, John Chan, Sharon Pifko-Hirst, Stephen A. Udem, Martin A. Billeter

Research output: Contribution to journalArticle

14 Citations (Scopus)

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)379-384
Number of pages6
JournalEMBO Journal
Volume9
Issue number2
StatePublished - 1990

Fingerprint

Measles virus
Viruses
Complementary DNA
RNA Viruses
Genome
Transcription
DNA
Bacteriophage T3
Plasmids
Bacteriophages
Genes
Ribonucleosides
Bacteriophage T7
RNA
Microinjections
Retroviridae
Helper-Inducer T-Lymphocytes
Reverse Transcription
Cytoplasm
Amplification

Keywords

  • Genome reconstruction
  • Infectious cDNA
  • Measles virus
  • Negative strand RNA
  • RNA virus

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Ballart, I., Eschle, D., Cattaneo, R., Schmid, A., Metzler, M., Chan, J., ... Billeter, M. A. (1990). Infectious measles virus from cloned cDNA. EMBO Journal, 9(2), 379-384.

Infectious measles virus from cloned cDNA. / Ballart, Isidro; Eschle, Daniel; Cattaneo, Roberto; Schmid, Anita; Metzler, Martina; Chan, John; Pifko-Hirst, Sharon; Udem, Stephen A.; Billeter, Martin A.

In: EMBO Journal, Vol. 9, No. 2, 1990, p. 379-384.

Research output: Contribution to journalArticle

Ballart, I, Eschle, D, Cattaneo, R, Schmid, A, Metzler, M, Chan, J, Pifko-Hirst, S, Udem, SA & Billeter, MA 1990, 'Infectious measles virus from cloned cDNA', EMBO Journal, vol. 9, no. 2, pp. 379-384.
Ballart I, Eschle D, Cattaneo R, Schmid A, Metzler M, Chan J et al. Infectious measles virus from cloned cDNA. EMBO Journal. 1990;9(2):379-384.
Ballart, Isidro ; Eschle, Daniel ; Cattaneo, Roberto ; Schmid, Anita ; Metzler, Martina ; Chan, John ; Pifko-Hirst, Sharon ; Udem, Stephen A. ; Billeter, Martin A. / Infectious measles virus from cloned cDNA. In: EMBO Journal. 1990 ; Vol. 9, No. 2. pp. 379-384.
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