Quantitative assays for maedi-visna virus genetic sequences and mRNA's based on RT-PCR with real-time FRET measurements.

2.50
Hdl Handle:
http://hdl.handle.net/2336/20859
Title:
Quantitative assays for maedi-visna virus genetic sequences and mRNA's based on RT-PCR with real-time FRET measurements.
Authors:
Gudmundsson, Bjarki; Bjarnadottir, Helga; Kristjansdottir, Steinunn; Jonsson, Jon J
Citation:
Virology 2003, 307(1):135-42
Issue Date:
1-Mar-2003
Abstract:
We developed robust, ultrasensitive, and accurate quantitative assays for maedi-visna virus (MVV) RNA and DNA genomic sequences and mRNA's expressed at various stages of lentiviral replication. Assay design was based on PCR with real-time fluorescence resonance energy transfer measurements. Specific assays were developed for gag-pol (genomic), tat, rev, env, and vif transcripts. Assay linearity ranged from 60 to 6 x 10(7) copies of target DNA. All assays were able to detect and measure corresponding mRNA's in MVV-infected FOS cells, whereas no signal was detected in mock-treated cells. In addition, RT-PCR based on amplification of gag sequences could be used to quantify RNA genomic sequences in supernatants from infected cells. These quantitative assays can be used to study the role of genetic elements in MVV infection and pathogenesis. They also allow rapid testing of lentiviral vectors and packaging systems based on MVV.
Description:
To access publisher full text version of this article. Please click on the hyperlink in Additional Links field
Additional Links:
http://www.sciencedirect.com/science/article/B6WXR-481FRSS-9/2/2c23143ad517d3d3884abc2b5aeeb132

Full metadata record

DC FieldValue Language
dc.contributor.authorGudmundsson, Bjarki-
dc.contributor.authorBjarnadottir, Helga-
dc.contributor.authorKristjansdottir, Steinunn-
dc.contributor.authorJonsson, Jon J-
dc.date.accessioned2008-03-17T15:08:38Z-
dc.date.available2008-03-17T15:08:38Z-
dc.date.issued2003-03-01-
dc.date.submitted2008-03-17-
dc.identifier.citationVirology 2003, 307(1):135-42en
dc.identifier.issn0042-6822-
dc.identifier.pmid12667821-
dc.identifier.urihttp://hdl.handle.net/2336/20859-
dc.descriptionTo access publisher full text version of this article. Please click on the hyperlink in Additional Links fielden
dc.description.abstractWe developed robust, ultrasensitive, and accurate quantitative assays for maedi-visna virus (MVV) RNA and DNA genomic sequences and mRNA's expressed at various stages of lentiviral replication. Assay design was based on PCR with real-time fluorescence resonance energy transfer measurements. Specific assays were developed for gag-pol (genomic), tat, rev, env, and vif transcripts. Assay linearity ranged from 60 to 6 x 10(7) copies of target DNA. All assays were able to detect and measure corresponding mRNA's in MVV-infected FOS cells, whereas no signal was detected in mock-treated cells. In addition, RT-PCR based on amplification of gag sequences could be used to quantify RNA genomic sequences in supernatants from infected cells. These quantitative assays can be used to study the role of genetic elements in MVV infection and pathogenesis. They also allow rapid testing of lentiviral vectors and packaging systems based on MVV.en
dc.language.isoenen
dc.publisherAcademic Pressen
dc.relation.urlhttp://www.sciencedirect.com/science/article/B6WXR-481FRSS-9/2/2c23143ad517d3d3884abc2b5aeeb132en
dc.subject.meshAnimalsen
dc.subject.meshBase Sequenceen
dc.subject.meshCell Lineen
dc.subject.meshDNA Primersen
dc.subject.meshFluorescence Resonance Energy Transferen
dc.subject.meshMacrophagesen
dc.subject.meshPolymerase Chain Reactionen
dc.subject.meshRNA, Messengeren
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen
dc.subject.meshSheepen
dc.subject.meshVisna-maedi virusen
dc.titleQuantitative assays for maedi-visna virus genetic sequences and mRNA's based on RT-PCR with real-time FRET measurements.en
dc.typeArticleen
dc.contributor.departmentDepartment of Biochemistry and Molecular Biology, University of Iceland Faculty of Medicine, Reykjavik, Iceland.en
dc.identifier.journalVirologyen

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