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Cited by 204

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Infectious cDNA Clone of the Epidemic West Nile Virus from New York City

Open access
Shi, Pei-Yong, Tilgner, Mark, Lo, Michael K., Kent, Kim A., Bernard, Kristen A.·2002·Journal of Virology

Abstract

We report the first full-length infectious clone of the current epidemic, lineage I, strain of West Nile virus (WNV). The full-length cDNA was constructed from reverse transcription-PCR products of viral RNA from an isolate collected during the year 2000 outbreak in New York City. It was cloned into plasmid pBR322 under the control of a T7 promoter and stably amplified in Escherichia coli HB101. RNA transcribed from the full-length cDNA clone was highly infectious upon transfection into BHK-21 cells, resulting in progeny virus with titers of 1 × 10 9 to 5 × 10 9 PFU/ml. The cDNA clone was engineered to contain three silent nucleotide changes to create a Sty I site (C to A and A to G at nucleotides [nt] 8859 and 8862, respectively) and to knock out an Eco RI site (A to G at nt 8880). These genetic markers were retained in the recovered progeny virus. Deletion of the 3′-terminal 199 nt of the cDNA transcript abolished the infectivity of the RNA. The plaque morphology, in vitro growth characteristics in mammalian and insect cells, and virulence in adult mice were indistinguishable for the parental and recombinant viruses. The stable infectious cDNA clone of the epidemic lineage I strain will provide a valuable experimental system to study the pathogenesis and replication of WNV.

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Metadata

Authors
Shi, P.-Y., Tilgner, M., Lo, M. K., Kent, K. A., & Bernard, K. A.
Venue
Journal of Virology
Published
2002-06-15
Volume
76
Issue
12
Pages
5847-5856
ISSN
  • 0022-538X (ISSN-L)
  • 1098-5514
OpenAlex
W2171529151
Cited by
204
References
1
FWCI
8.66
Type
journal article
Language
en
Open access
green

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CoChat. (2026). Citation check: Infectious cDNA Clone of the Epidemic West Nile Virus from New York City. https://app.cochat.ai/papers/10.1128/jvi.76.12.5847-5856.2002
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Shi, P.-Y., Tilgner, M., Lo, M. K., Kent, K. A., & Bernard, K. A. (2002). Infectious cDNA Clone of the Epidemic West Nile Virus from New York City. Journal of Virology. https://doi.org/10.1128/jvi.76.12.5847-5856.2002

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