Successful application of FTA® Classic Card technology and use of bacteriophage 29 DNA polymerase for large-scale field sampling and cloning of complete maize streak virus genomes

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dc.contributor.authorOwor, Betty E.
dc.contributor.authorShepherd, Dionne N.
dc.contributor.authorTaylor, Nigel J.
dc.contributor.authorEdema, Richard
dc.contributor.authorMonjane, Ad´erito L.
dc.contributor.authorThomson, Jennifer A.
dc.contributor.authorMartin, Darren P.
dc.contributor.authorVarsani, Arvind
dc.date.accessioned2022-05-26T20:46:11Z
dc.date.available2022-05-26T20:46:11Z
dc.date.issued2007
dc.description.abstractLeaf samples from 155 maize streak virus (MSV)-infected maize plants were collected from 155 farmers’ fields in 23 districts in Uganda in May/June 2005 by leaf-pressing infected samples onto FTA® Classic Cards. Viral DNA was successfully extracted from cards stored at room temperature for 9 months. The diversity of 127 MSV isolates was analysed by PCR-generated RFLPs. Six representative isolates having different RFLP patterns and causing either severe, moderate or mild disease symptoms, were chosen for amplification from FTA cards by bacteriophage 29 DNA polymerase using the TempliPhiTM system. Full-length genomes were inserted into a cloning vector using a unique restriction enzyme site, and sequenced. The 1.3-kb PCR product amplified directly from FTA-eluted DNA and used for RFLP analysis was also cloned and sequenced. Comparison of cloned whole genome sequences with those of the original PCR products indicated that the correct virus genome had been cloned and that no errors were introduced by the 29 polymerase. This is the first successful large-scale application of FTA card technology to the field, and illustrates the ease with which large numbers of infected samples can be collected and stored for downstream molecular applications such as diversity analysis and cloning of potentially new virus genomes. © 2006 Elsevier B.V. All rights reserved.en_US
dc.identifier.citationOwor, BE, Shepherd, DN, Taylor, NJ, Edema, R., Monjane, AL, Thomson, JA, ... & Varsani, A. (2007). Successful application of FTA® Classic Card technology and use of bacteriophage ϕ29 DNA polymerase for large-scale field sampling and cloning of complete maize streak virus genomes. Journal of virological methods , 140 (1-2), 100-105. doi:10.1016/j.jviromet.2006.11.004en_US
dc.identifier.other10.1016/j.jviromet.2006.11.004
dc.identifier.urihttps://nru.uncst.go.ug/handle/123456789/3523
dc.language.isoenen_US
dc.publisherJournal of virological methodsen_US
dc.subjectMaize streak virus (MSV)en_US
dc.subjectGeminivirusen_US
dc.subjectFTA cardsen_US
dc.subjectRolling circle amplificationen_US
dc.subjectPhi29 DNA polymeraseen_US
dc.subjectMSV diversityen_US
dc.titleSuccessful application of FTA® Classic Card technology and use of bacteriophage 29 DNA polymerase for large-scale field sampling and cloning of complete maize streak virus genomesen_US
dc.typeArticleen_US
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