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Molecular Plant Pathology

Molecular Cloning and Physical Mapping of Potato Virus S Complementary DNA. J. Monis, Graduate student, Department of Plant Pathology, University of Wisconsin-Madison, Madison 53706, Present address: Department of Pharmacology, Southwestern Medical Center, University of Texas, Dallas 75235; G. A. de Zoeten, professor, Department of Plant Pathology, University of Wisconsin-Madison, Madison 53706, Present address: Department of Botany and Plant Pathology, University of Michigan, East Lansing 48823. Phytopathology 80:446-450. Accepted for publication 13 September 1989. Copyright 1990 The American Phytopathological Society. DOI: 10.1094/Phyto-80-446.

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.