Plant Molecular Biology

, Volume 6, Issue 6, pp 455–465 | Cite as

An analysis of tobacco mosaic virus replicative structures synthesized in vitro

  • Nevin Dale Young
  • Milton Zaitlin


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.


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


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Copyright information

© Martinus Nijhoff Publishers 1986

Authors and Affiliations

  • Nevin Dale Young
    • 1
  • Milton Zaitlin
    • 1
  1. 1.Department of Plant PathologyCornell UniversityIthacaU.S.A.

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