An analysis of tobacco mosaic virus replicative structures synthesized in vitro

Nevin Dale Young, Milton Zaitlin

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18 Scopus citations


The RNA structures synthesized in vitro by a crude enzyme complex from tobacco mosaic virus (TMV)-infected leaves have been analyzed; the major viral-specific products were similar to TMV-replicative form (RF) and-replicative intermediate (RI) in electrophoretic behavior and ribonuclease sensitivity. Synthesis of these RF-like and RI-like structures neither required nor responded to added viral RNA, but did require all four ribonucleotide triphosphates. Enriched radiolabeled RF-like and RI-like RNA fractions were isolated from non-denaturing agarose gels by electroelution and hybridized to a collection of TMV sequences cloned into bacteriophage M13. Enriched RF-RNA hybridized to sequences of both plus and minus polarity, while enriched RI-RNA hybridized only to inserts of minus polarity, indicating only plus strand synthesis in this fraction. Most of the label incorporated into the plus strand of the enriched RF-RNA was found near the 3′-end of this strand, while most of the label incorporated into enriched RI-RNA was found several hundred bases from the 5′-end of the plus strand.

Original languageEnglish (US)
Pages (from-to)455-465
Number of pages11
JournalPlant molecular biology
Issue number6
StatePublished - Nov 1986


  • molecular hybridization
  • plant viral replication
  • replicative form
  • replicative intermediate
  • tobacco mosaic virus


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