Journal
JOURNAL OF GENERAL VIROLOGY
Volume 93, Issue -, Pages 1991-1995Publisher
MICROBIOLOGY SOC
DOI: 10.1099/vir.0.043752-0
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Funding
- German Ministry of Education (project on Chikungunya virus blood donor screening) [01ES0814]
- European Commission contract ArboZooNet [211757]
- European Commission (contract EVA) [228292]
- European Commission (contract EMPERIE) [223498]
- European Commission (contract ICRES) [261202]
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Chikungunya virus (CHIKV) has caused massive epidemics in the Indian Ocean region since 2005. It belongs to the genus Alpha virus and possesses a positive-stranded RNA genome of nearly 12 kb in size. To produce genetically modified viruses for the study of various aspects of the CHIKV life cycle, a reverse genetic system is needed. We report the generation of a T7 RNA polymerase-driven infectious cDNA clone of CHIKV. Electroporation of in vitro-transcribed RNA resulted in the recovery of a recombinant virus with growth characteristics comparable to the parental strain. Using the established cDNA clone, the red fluorescent marker gene mCherry was introduced into two different sites within the CHIKV nsP3 gene. Both constructs allowed the rescue of stable fluorescent reporter viruses with growth characteristics similar to the wild-type virus. The latter reporter viruses represent valuable tools for easy follow-up of replicating CHIKV useful in several applications of CHIKV research.
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