Key Points
- Clone the full-length genome of cardiotropic Coxsackie B3 virus to establish an infectious, biologically active cDNA copy for molecular and diagnostic research.
- Synthesized full-length double-stranded cDNA of approximately 7,500 nucleotides from the single-stranded RNA genome of Coxsackie B3 virus (Nancy strain).
- Transfected recombinant plasmid DNA into mammalian cell cultures and characterized the clone using restriction enzyme digestion and partial 5' end nucleotide sequencing.
- Transfection of the cloned full-length recombinant cDNA successfully yielded infectious virus particles that were antigenically identical to wild-type Coxsackie B3 virus.
- Restriction enzyme mapping and partial nucleotide sequencing confirmed the intact structural identity of the approximately 7,500-nucleotide biologically active viral cDNA clone.
Structured PICO
PPopulationMammalian cells and single-stranded RNA genome of the cardiotropic Coxsackie B3 virus (Nancy strain)
IInterventionTransfection of recombinant plasmid DNA containing full-length reverse-transcribed cloned viral cDNA (approximately 7500 nucleotides)
OOutcomeGeneration of infectious antigenically identical Coxsackie B3 virussurrogate
The successful molecular cloning of a biologically active viral cDNA copy of the cardiotropic Coxsackie B3 virus provides a tool to study enteroviral heart disease and a potential diagnostic means.