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Plant Mol Biol. 1986 Nov;6(6):455-65. doi: 10.1007/BF00027137.

An analysis of tobacco mosaic virus replicative structures synthesized in vitro.

Author information

1
Department of Plant Pathology, Cornell University, Ithaca, NY, 14853, U.S.A..

Abstract

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.

PMID:
24307422
DOI:
10.1007/BF00027137

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