Construction of infectious cDNA clones for RNA viruses: Turnip crinkle virus.
Ryabov, Eugene V. (2008) Construction of infectious cDNA clones for RNA viruses: Turnip crinkle virus. In: Plant virology protocols: from virus sequence to protein function. Methods in Molecular Biology, Vol.451 . Totowa, N.J: Humana Press, Inc., pp. 491-502. ISBN 978-1-58829-827-0 ISSN: 1064-3745Full text not available from this repository.
Official URL: http://dx.doi.org/10.1007/978-1-59745-102-4_33
Reverse genetic approach is widely used in virology as it makes possible direct identification of viral gene function and uses RNA genomes as vectors. Production of infectious cDNA clones is an essential step in developing a reverse genetic system for an RNA virus. Here, we present rapid method for generation of infectious cDNA clone for Turnip crinkle virus (TCV). The infectious cDNA clone could be used for production of in vitro transcripts with the T7 RNA polymerase which could be used for infection of plants or plant cell protoplasts. The procedure described here includes purification of TCV, viral RNA extraction, reverse transcription, PCR amplification of the full-length cDNA copy of TCV linked to a T7 RNA polymerase promoter, cloning into a plasmid vector, in vitro transcription, and selection of infectious clones.
|Item Type:||Book Item|
|Subjects:||Q Science > Q Science (General)|
|Divisions:||Faculty of Science > Life Sciences (2010- )|
|Series Name:||Methods in Molecular Biology|
|Journal or Publication Title:||Methods in molecular biology (Clifton, N.J.)|
|Publisher:||Humana Press, Inc.|
|Place of Publication:||Totowa, N.J|
|ISBN:||978-1-58829-827-0 ISSN: 1064-3745|
|Book Title:||Plant virology protocols: from virus sequence to protein function|
|Number of Pages:||12|
|Page Range:||pp. 491-502|
|Status:||Not Peer Reviewed|
|Access rights to Published version:||Restricted or Subscription Access|
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