Molecular Cloning and Physical Mapping of Potato Virus S Complementary DNA

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Abstract
Complementary DNA clones that represent almost the full length of the Andean strain of potato virus S (PVS-An) genomic RNA were produced. Complementary DNA of 7.5 kilobase pairs (kbp) was synthesized and was either digested with restriction enzymes to produce cohesive ends or attached to EcoRI linkers and cloned into Escherichia coli plasmids pUC19 and pTZ18R. The size of the overlapping clones ranged from 0.4 to 6.0 kbp. A physical map of the overlapping clones was produced by restriction endonuclease digestion and Southern blot hybridization analysis. The clones have been successfully used in northern and dot blot hybridization assays to detect PVS-An specific sequences in crude leaf extracts of potato and Chenopodium quinoa infected with this virus. Furthermore, the sequence of the 5'' end of PVS-An not represented in the cDNA (261 bases) was obtained.