Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay

Pan Tao Zhang, Chao Shan, Xiao Dan Li, Si Qing Liu, Cheng Lin Deng, Han Qing Ye, Bao Di Shang, Pei-Yong Shi, Ming Lv, Bei Fen Shen, Cheng Feng Qin, Bo Zhang

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

West Nile virus (WNV) is a neurotropic human pathogen that has caused increasing infected cases over recent years. There is currently no licensed vaccine or effective drug for prevention and treatment of WNV infection in humans. To facilitate antiviral drug discovery and neutralizing antibody detection, a WNV cDNA clone containing a luciferase reporter gene was constructed through incorporating Gaussia luciferase (Gluc) gene within the capsid-coding region of WNV genome. Transfection of BHK-21 cells with the cDNA clone-derived RNA generated luciferase reporter WNV (WNV-Gluc) and the stable WNV-Gluc with high titers (>107PFU/ml) was obtained through plaque purification. Luciferase activity was used to effectively quantify the viral production of WNV-Gluc. Using the reporter virus WNV-Gluc, we developed a luciferase based assay in a 12-well format for evaluating neutralizing antibodies. The reporter virus could be a powerful tool for epidemiological investigation of WNV, vaccine evaluation, antiviral drug screening, and the study of WNV replication and pathogenesis.

Original languageEnglish (US)
Pages (from-to)17-24
Number of pages8
JournalVirus Research
Volume211
DOIs
StatePublished - Jan 4 2016
Externally publishedYes

Fingerprint

West Nile virus
Luciferases
Neutralizing Antibodies
Antiviral Agents
West Nile Virus Vaccines
Complementary DNA
Clone Cells
Viruses
Preclinical Drug Evaluations
Capsid
Virus Diseases
Drug Discovery
Virus Replication
Reporter Genes
Transfection
Vaccines
Genome
RNA

Keywords

  • Antiviral drug discovery
  • Gaussia luciferase
  • Neutralizing antibody
  • Reporter virus
  • West nile virus

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases
  • Cancer Research

Cite this

Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay. / Zhang, Pan Tao; Shan, Chao; Li, Xiao Dan; Liu, Si Qing; Deng, Cheng Lin; Ye, Han Qing; Shang, Bao Di; Shi, Pei-Yong; Lv, Ming; Shen, Bei Fen; Qin, Cheng Feng; Zhang, Bo.

In: Virus Research, Vol. 211, 04.01.2016, p. 17-24.

Research output: Contribution to journalArticle

Zhang, PT, Shan, C, Li, XD, Liu, SQ, Deng, CL, Ye, HQ, Shang, BD, Shi, P-Y, Lv, M, Shen, BF, Qin, CF & Zhang, B 2016, 'Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay', Virus Research, vol. 211, pp. 17-24. https://doi.org/10.1016/j.virusres.2015.09.015
Zhang, Pan Tao ; Shan, Chao ; Li, Xiao Dan ; Liu, Si Qing ; Deng, Cheng Lin ; Ye, Han Qing ; Shang, Bao Di ; Shi, Pei-Yong ; Lv, Ming ; Shen, Bei Fen ; Qin, Cheng Feng ; Zhang, Bo. / Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay. In: Virus Research. 2016 ; Vol. 211. pp. 17-24.
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