Construction of Infectious Clones for RNA Viruses: TMV
- 1 January 2008
- book chapter
- Published by Springer Science and Business Media LLC in Methods in molecular biology (Clifton, N.J.)
- Vol. 451, 477-490
- https://doi.org/10.1007/978-1-59745-102-4_32
Abstract
The generation of infectious clones is routinely the first step for reverse genetic studies of RNA plant virus gene and sequence function. The procedure given here, details the creation of cDNA clones of tobacco mosaic virus, from which infectious transcripts can be generated in vitro with T7 RNA polymerase. The procedure describes methods for virion purification, viral RNA extraction, reverse transcription, PCR amplification of genomic cDNA fragments, generation of a full-length cDNA clone under the control of a T7 promoter, in vitro transcription, and infectivity testing.This publication has 19 references indexed in Scilit:
- Odontoglossum ringspot virus host range restriction in Nicotiana sylvestris maps to the replicase geneMolecular Plant Pathology, 2005
- Systemic Agrobacterium tumefaciens–mediated transfection of viral replicons for efficient transient expression in plantsNature Biotechnology, 2005
- Improvement of the movement and host range properties of a plant virus vector through DNA shufflingThe Plant Journal, 2002
- Deletion of Internal Sequences Results in Tobacco Mosaic Virus Defective RNAs That Accumulate to High Levels without Interfering with Replication of the Helper VirusVirology, 1998
- A CaMV 35S promoter driven cDNA clone of tobacco mosaic virus can infect host plant tissue despite being uninfectious when manually inoculated onto leavesArchiv für die gesamte Virusforschung, 1997
- Mapping Nucleotides in the 126-kDa Protein Gene That Control the Differential Symptoms Induced by Two Strains of Tobacco Mosaic VirusVirology, 1996
- Transfection of whole plants from wounds inoculated with Agrobacterium tumefaciens containing cDNA of tobacco mosaic virusJournal of Virological Methods, 1993
- In vivo complementation of infectious transcripts from mutant tobacco mosaic virus cDNAs in transgenic plantsVirology, 1991
- DNA Probes: Applications of the Principles of Nucleic Acid HybridizationCritical Reviews in Biochemistry and Molecular Biology, 1991
- Simple RNA enzymes with new and highly specific endoribonuclease activitiesNature, 1988