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