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Volume 80, Issue 12

Characterization of infectious Murray Valley encephalitis virus derived from a stably cloned genome-length cDNA Free

Abstract

An infectious cDNA clone of Murray Valley encephalitis virus prototype strain 1-51 (MVE-1-51) was constructed by stably inserting genome-length cDNA into the low-copy-number plasmid vector pMC18. Designated pMVE-1-51, the clone consisted of genome-length cDNA of MVE-1-51 under the control of a T7 RNA polymerase promoter. The clone was constructed by using existing components of a cDNA library, in addition to cDNA of the 3′ terminus derived by RT–PCR of poly(A)-tailed viral RNA. Upon comparison with other flavivirus sequences, the previously undetermined sequence of the 3′ UTR was found to contain elements conserved throughout the genus . RNA transcribed from pMVE-1-51 and subsequently transfected into BHK-21 cells generated infectious virus. The plaque morphology, replication kinetics and antigenic profile of clone-derived virus (CDV-1-51) was similar to the parental virus . Furthermore, the virulence properties of CDV-1-51 and MVE-1-51 (LD values and mortality profiles) were found to be identical in the mouse model. Through site-directed mutagenesis, the infectious clone should serve as a valuable tool for investigating the molecular determinants of virulence in MVE virus.

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© Microbiology Society
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1999-12-01
2024-04-16
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Characterization of infectious Murray Valley encephalitis virus derived from a stably cloned genome-length cDNA
J. Gen. Virol. 80, 3115 (1999); https://doi.org/10.1099/0022-1317-80-12-3115
/content/journal/jgv/10.1099/0022-1317-80-12-3115
/content/journal/jgv/10.1099/0022-1317-80-12-3115
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