Synthesis of biologically active cDNA clones of cymbidium mosaic potexvirus using a population cloning strategy

Biologically active cDNA clones of cymbidium mosaic potexvirus (CymMV) were synthesized using a population cloning strategy. Three populations of overlapping RT-PCR products encompassing the entire viral RNA of CymMV were ligated into pBluescriptKS+ with T7 RNA polymerase promoter fused to the 5' extreme of the viral cDNA. Capped-RNA in vitro transcripts were infectious. This is the first report of successful synthesis of biologically active CymMV clones. Unlike the conventional methods, population cloning maximizes the probability of obtaining biologically active cDNA clones.