Recovery of a chemically synthesized Japanese encephalitis virus reveals two critical adaptive mutations in NS2B and NS4A

Xiao Dan Li, Xiao Feng Li, Han Qing Ye, Cheng Lin Deng, Qing Ye, Chao Shan, Bao Di Shang, Lin Lin Xu, Shi Hua Li, Sheng Bo Cao, Zhi Ming Yuan, Pei-Yong Shi, Cheng Feng Qin, Bo Zhang

Research output: Contribution to journalArticle

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Abstract

A full-length genome infectious clone is a powerful tool for functional assays in virology. In this study, using a chemical synthesized complete genome of Japanese encephalitis virus (JEV) strain SA14 (GenBank accession no. U14163), we constructed a full-length genomic cDNA clone of JEV. The recovered virus from the cDNA clone replicated poorly in baby hamster kidney (BHK-21) cells and in suckling mice brain. Following serial passage in BHK-21 cells, adaptive mutations within the NS2B and NS4A proteins were recovered in the passaged viruses leading to viruses with a large-plaque phenotype. Mutagenesis analysis, using a genome-length RNA and a replicon of JEV, demonstrated that the adaptive mutations restored replication to different degrees, and the restoration efficiencies were in the order: NS2B-T102M

Original languageEnglish (US)
Pages (from-to)806-815
Number of pages10
JournalJournal of General Virology
Volume95
Issue numberPART 4
DOIs
StatePublished - 2014
Externally publishedYes

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Japanese Encephalitis Virus
Clone Cells
Genome
Viruses
Mutation
Complementary DNA
Serial Passage
Replicon
Virology
Nucleic Acid Databases
Mutagenesis
Cricetinae
RNA
Phenotype
Kidney
Brain
Proteins

ASJC Scopus subject areas

  • Virology

Cite this

Recovery of a chemically synthesized Japanese encephalitis virus reveals two critical adaptive mutations in NS2B and NS4A. / Li, Xiao Dan; Li, Xiao Feng; Ye, Han Qing; Deng, Cheng Lin; Ye, Qing; Shan, Chao; Shang, Bao Di; Xu, Lin Lin; Li, Shi Hua; Cao, Sheng Bo; Yuan, Zhi Ming; Shi, Pei-Yong; Qin, Cheng Feng; Zhang, Bo.

In: Journal of General Virology, Vol. 95, No. PART 4, 2014, p. 806-815.

Research output: Contribution to journalArticle

Li, XD, Li, XF, Ye, HQ, Deng, CL, Ye, Q, Shan, C, Shang, BD, Xu, LL, Li, SH, Cao, SB, Yuan, ZM, Shi, P-Y, Qin, CF & Zhang, B 2014, 'Recovery of a chemically synthesized Japanese encephalitis virus reveals two critical adaptive mutations in NS2B and NS4A', Journal of General Virology, vol. 95, no. PART 4, pp. 806-815. https://doi.org/10.1099/vir.0.061838-0
Li, Xiao Dan ; Li, Xiao Feng ; Ye, Han Qing ; Deng, Cheng Lin ; Ye, Qing ; Shan, Chao ; Shang, Bao Di ; Xu, Lin Lin ; Li, Shi Hua ; Cao, Sheng Bo ; Yuan, Zhi Ming ; Shi, Pei-Yong ; Qin, Cheng Feng ; Zhang, Bo. / Recovery of a chemically synthesized Japanese encephalitis virus reveals two critical adaptive mutations in NS2B and NS4A. In: Journal of General Virology. 2014 ; Vol. 95, No. PART 4. pp. 806-815.
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