A novel bacterium-free method for generation of flavivirus infectious DNA by circular polymerase extension reaction allows accurate recapitulation of viral heterogeneity

Judith Edmonds, Erinke van Grinsven, Natalie Prow, Angela Bosco-Lauth, Aaron Brault, Richard A. Bowen, Roy A. Hall, Alexander A. Khromykh

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

A novel bacterium-free approach for rapid assembly of flavivirus infectious cDNAs using circular polymerase extension reaction was applied to generate infectious cDNA for the virulent New South Wales isolate of the Kunjin strain of West Nile virus (KUNV) that recently emerged in Australia. Recovered virus recapitulated the genetic heterogeneity present in the original isolate. The approach was utilized to generate viral mutants with designed phenotypic properties and to identify E protein glycosylation as one of the virulence determinants.

Original languageEnglish (US)
Pages (from-to)2367-2372
Number of pages6
JournalJournal of Virology
Volume87
Issue number4
DOIs
StatePublished - Feb 1 2013
Externally publishedYes

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Flavivirus
West Nile virus
DNA-Directed DNA Polymerase
glycosylation
New South Wales
virulence
Complementary DNA
Bacteria
viruses
mutants
Genetic Heterogeneity
bacteria
DNA
Glycosylation
Virulence
proteins
Viruses
methodology

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

A novel bacterium-free method for generation of flavivirus infectious DNA by circular polymerase extension reaction allows accurate recapitulation of viral heterogeneity. / Edmonds, Judith; van Grinsven, Erinke; Prow, Natalie; Bosco-Lauth, Angela; Brault, Aaron; Bowen, Richard A.; Hall, Roy A.; Khromykh, Alexander A.

In: Journal of Virology, Vol. 87, No. 4, 01.02.2013, p. 2367-2372.

Research output: Contribution to journalArticle

Edmonds, Judith ; van Grinsven, Erinke ; Prow, Natalie ; Bosco-Lauth, Angela ; Brault, Aaron ; Bowen, Richard A. ; Hall, Roy A. ; Khromykh, Alexander A. / A novel bacterium-free method for generation of flavivirus infectious DNA by circular polymerase extension reaction allows accurate recapitulation of viral heterogeneity. In: Journal of Virology. 2013 ; Vol. 87, No. 4. pp. 2367-2372.
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