Molecular Cloning of Single-stranded RNAs of Potato Leafroll Virus and Beet Western Yellows Virus

Abstract
Complementary DNA libraries were synthesized using the genomic RNAs of potato leafroll virus (PLRV) and beet western yellows virus (BWYV), by random as well as oligo(dT) priming of polyadenylated RNA, and cloned into pBR322. Two restriction endonuclease maps were constructed, extending to 61 kbp and 5.5 kbp for the PLRV and BWYV genomes respectively. The 3'' ends of genomic RNAs were verified by sequence analysis of oligo(dT)-primed clones; 5''-terminal sequences of PLRV snd BWYV were not detected among the cDNA fragments mapped. Selected clones were used to demonstrate specific virus detection in total nucleic acid preparations of PLRV- or BWYV-infected Physalis floridana Rydb. One PLRV-derived clone showed cross-hybridization with purified BWYV RNA.