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Reverse Genetic Analysis of Adaptive Mutations within the Capsid Proteins of Enterovirus 71 (EV-A71) Strains Necessary for Infection of CHO-K1 Cells

  • Carla Bianca Luena Victorio
  • Yishi Xu
  • Qimei Ng
  • Vincent T. K. Chow
  • Kaw Bing ChuaEmail author
Letter
  • 106 Downloads

Dear Editor,

Previous studies had described the adaptation of enterovirus 71 (EV-A71) strains that enabled entry and viral replication in Chinese Hamster Ovary (CHO) cell line (Zaini and McMinn 2012; Zaini et al.2012). These adapted strains derived from serial passage of a clinical isolate in CHO cells exhibited an amino acid substitution at VP2149, which enhanced viral replication by 100~1000-fold compared to the clinical isolate. The VP2149 mutation was claimed responsible for adaptation to CHO-K1 cells without performing detailed molecular analyses to support these claims. In this study, we evaluate various VP1 and VP2 mutations in two CHO-adapted EV-A71 strains derived in our lab to assess their contribution to the phenotype of CHO cell adaptation.

Two EV-A71 strains derived in our laboratory and found to productively infect CHO cells, EV71:TLLm (Genbank accession no. KF514879) and EV71:TLLcho (Genbank accession no. KM508794.1), were evaluated in this study. The adaptation history...

Notes

Acknowledgements

This research was fully funded by Temasek Lifesciences Laboratory Ltd. The pCMV-T7pol plasmid construct was a generous gift by Dr. Peter McMinn from Sydney University, Western Australia.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Animal and Human Rights Statement

This article does not contain any studies with human or animal subjects performed by any of the authors.

Supplementary material

12250_2019_167_MOESM1_ESM.pdf (6.3 mb)
Supplementary material 1 (PDF 6432 kb)

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Copyright information

© Wuhan Institute of Virology, CAS 2019

Authors and Affiliations

  1. 1.Temasek Lifesciences Laboratory, 1 Research LinkNational University of SingaporeSingaporeSingapore
  2. 2.Host and Pathogen Interactivity Laboratory, Department of Microbiology, Yong Loo Lin School of Medicine, National University Health SystemNational University of SingaporeSingaporeSingapore
  3. 3.Laboratory for Translational and Molecular Imaging, Cancer and Stem Cell Biology ProgrammeDuke-NUS Medical SchoolSingaporeSingapore

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