Gene Mapping and Expression of Tomato Bushy Stunt Virus

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Abstract
Cloned DNA species complementary to the RNA of tomato bushy stunt virus (TBSV) were produced and the location of each on the virus genome was determined. Polysomes were prepared from TBSV-infected plants and RNA, released from the polysomes by treatment with puromycin, was shown by Northern hybridization to contain TBSV genomic RNA (4.8 kb) and two subgenomic ssRNA species of 2.0 and 0.9 kgb. Hybrid selection using clones corresponding to different parts of the virus genome showed that the two subgenomic RNA species are probably 3'' coterminal with the genomic RNA. In vitro translation of hybrid-selected RNA showed that the genomic (4.8 kb), 2.0 and 0.9 kb RNA species encode proteins of Mr 37 000 (37K), 40K (the virus coat protein) and 22K respectively. Translation in the presence of calf liver tRNA produced, in addition to the other products, a protein of Mr 90K which may be a readthrough product of the 37K protein. The results have enabled a model for the genome organization and expression of TBSV to be formulated.