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: 1064-3745

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Abstract

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
ISSN:	1064-3745
Book Title:	Plant virology protocols: from virus sequence to protein function
Date:	2008
Volume:	Vol.451
Number of Pages:	12
Page Range:	pp. 491-502
Identification Number:	10.1007/978-1-59745-102-4_33
Status:	Not Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
URI:	http://wrap.warwick.ac.uk/id/eprint/42821

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