Development of a stable Gaussia luciferase enterovirus 71 reporter virus

Lin Lin Xu, Chao Shan, Cheng Lin Deng, Xiao Dan Li, Bao Di Shang, Han Qing Ye, Si Qing Liu, Zhi Ming Yuan, Qing Yin Wang, Pei Yong Shi, Bo Zhang

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

We report a stable Gaussia luciferase enterovirus 71 (Gluc-EV71) reporter virus to facilitate drug discovery. The Gluc-EV71 reporter virus was generated by engineering the Gaussia luciferase (Gluc) gene between the 5' untranslated region and VP4 gene of the EV71 genome. We could recover Gluc-EV71 after transfection of Vero cells with the cDNA clone-derived RNA. The reporter virus efficiently infects and replicates in various cell types (Vero, human rhabdomyosarcoma, and HeLa cells), producing robust luciferase activity. The Gluc-EV71 virus replicates slower than the wild-type virus in cell culture. The reporter virus is stable in maintaining the Gluc gene after five rounds of continuous passaging in Vero cells. Using known EV71 inhibitors, we demonstrate that the reporter virus can be used for antiviral testing. However, the Gluc-EV71 infection assay cannot be adapted to a homogenous format for high throughput screen, mainly due to the secreted nature of the Gluc protein and the short half-life of the Gluc luminescence signal. The Gluc-EV71 and its infection assay could be useful for antiviral drug discovery as well as for studying EV71 replication and pathogenesis.

Original languageEnglish (US)
Pages (from-to)62-66
Number of pages5
JournalJournal of Virological Methods
Volume219
DOIs
StatePublished - Jul 1 2015

Keywords

  • Antiviral
  • Enterovirus
  • Gaussia luciferase
  • Replication
  • Reporter virus

ASJC Scopus subject areas

  • Virology

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  • Cite this

    Xu, L. L., Shan, C., Deng, C. L., Li, X. D., Shang, B. D., Ye, H. Q., Liu, S. Q., Yuan, Z. M., Wang, Q. Y., Shi, P. Y., & Zhang, B. (2015). Development of a stable Gaussia luciferase enterovirus 71 reporter virus. Journal of Virological Methods, 219, 62-66. https://doi.org/10.1016/j.jviromet.2015.03.020