The nature of the RNA products synthesized in vitro with a cell-free extract from TMV-infected tobacco leaves.
A cell-free extract containing TMV-RNA replicase was prepared from TMV-infected tobacco leaves. It could synthesize double-stranded RNAs in the presence of four nucleoside triphosphates (among them, UTP was tritium-labelled), magnesium ion and actinomycin D. It was confirmed by polyacrylamide-agarose gel electrophoresis, RNase treatment, thermal denaturation and self annealing that 3H-ds RNAs, obtained from phenol-SDS extraction and Serva cellulose column chromatography, consisted of replicative form (RF) and replicative intermediate (RI) of TMV-RNA, with molecular weights of 40 X 10(6) and 5.0 X 10(6), respectively. Molecular hybridization competition experiment showed that 60-70% of the nascent RNAs in the 3H-ds RNA were plus strand of tMV-RNA.
D002474:Cell-Free System; D010947:Plants, Toxic; D012330:RNA, Double-Stranded / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis*; D014026:Nicotiana / Q000378:metabolism*; D014027:Tobacco Mosaic Virus / Q000378:metabolism*
D012330:RNA, Double-Stranded; D012367:RNA, Viral