title: Rescue of infective virus from a genome-length cDNA clone of the FMDV serotype O (IND-R2/75) vaccine strain and its characterization.
abstract: We report here the construction and characterization of an infectious cDNA clone of the Indian vaccine strain of foot and mouth disease virus (FMDV) serotype O, IND-R2/75. Viral genome was amplified by reverse transcription-polymerase chain reaction (RT-PCR) in five fragments and subsequently assembled sequentially in a plasmid vector to generate a complete cDNA clone, flanked by the T7 RNA polymerase promoter and poly (A) tail at 5' and 3' ends, respectively. Transfection of BHK-21 cells with the RNA transcribed from this genome-length cDNA construct allowed the recovery of infectious recombinant FMDV particles as evidenced by cytopathic effect in BHK-21 cells. Characterization of the recombinant virus revealed its similarity to the parental strain. Recombinant virus could be distinguished from the parental virus based on the presence of a unique marker sequence in the former, which was incorporated in the cDNA using a silent mutation. The virus showed no significant amino acid changes in the capsid-coding region when serially passaged up to ten times in BHK-21 cells, while retaining the marker sequence.
authors: Indian Veterinary Research Institute, Bellary Road, Hebbal, Bangalore 560 024, India.
authors: R
authors: R
authors: Rajasekhar
authors: Madhusudan
authors: M
authors: Hosamani
authors: Suresh H
authors: SH
authors: Basagoudanavar
authors: B P
authors: BP
authors: Sreenivasa
authors: R P
authors: RP
authors: Tamil Selvan
authors: Paramasivam
authors: P
authors: Saravanan
authors: Ramamurthy
authors: R
authors: Venkataramanan
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D005536:Foot-and-Mouth Disease / Q000517:prevention & control / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics* / Q000276:immunology; D016679:Genome, Viral; D007194:India; D009500:Neutralization Tests; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012703:Serotyping; D014162:Transfection; D014765:Viral Vaccines / Q000235:genetics* / Q000276:immunology
chemical_list: D012367:RNA, Viral; D014765:Viral Vaccines