title: Reverse Genetics Approaches for Chikungunya Virus.
abstract: Reverse genetic systems based on an infectious cDNA clone, a double-stranded copy of the viral genome carried on a plasmid vector, have greatly enhanced the understanding of RNA virus biology by facilitating genetic manipulation of viral RNA genomes. To date, infectious cDNA clones of Chikungunya virus (CHIKV) have been constructed using different combinations of plasmid vectors and/or bacterial host strains. Here, we describe our approaches for the construction of infectious cDNA clones of CHIKV and the protocol for genetic manipulation of the clones by site-directed mutagenesis.
authors.affiliation: Department of Pathobiology, Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen, Thailand. patphu@kku.ac.th.
authors.forename: Patchara
authors.initials: P
authors.lastname: Phuektes
authors.affiliation: Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
authors.forename: Justin Jang Hann
authors.initials: JJ
authors.lastname: Chu
mesh_terms: D000818:Animals; D002460:Cell Line; D002646:Chikungunya virus / Q000235:genetics*; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000235:genetics*; D059386:Reverse Genetics / Q000379:methods*; D014779:Virus Replication / Q000235:genetics
keywords: Chikungunya virus; Infectious cDNA clone; Reverse genetics; Site-directed mutagenesis