Title: A rapid and convenient variant of fusion-PCR to construct chimeric flaviviruses
Authors: Charlier, Nathalie
Molenkamp, Richard
Leyssen, Pieter
Vandamme, Anne-Mieke
De Clercq, Erik
Bredenbeek, Peter
Neyts, Johan # ×
Issue Date: Mar-2003
Series Title: Journal of virological methods vol:108 issue:1 pages:67-74
Abstract: So far, full-length cDNAs of chimeric flaviviruses have been constructed by restriction-enzyme cleavage of the gene(s) to be exchanged or by fusion-PCR of two amplified PCR fragments. The construction of a chimeric flavivirus by a faster and more convenient variant of the standard fusion-PCR is reported. A Modoc/yellow fever chimeric virus was engineered in which the structural prM and E genes of yellow fever virus 17D were replaced by the homologous genes of Modoc virus. In two PCR steps, a fusion was made between the 3' end of the C gene of yellow fever virus and the 5' end of the prM gene of Modoc virus, and between the 3' end of the E gene of Modoc virus and the 5' end of the NS1 gene of yellow fever virus. For each of the two fusions between yellow fever and Modoc virus, a standard PCR was performed to amplify a short fragment with one overlapping end that could be used as one of the primers in the subsequent (fusion) PCR.
ISSN: 0166-0934
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Laboratory of Virology and Chemotherapy (Rega Institute)
Faculty of Pharmaceutical Sciences - miscellaneous
Laboratory of Clinical and Epidemiological Virology (Rega Institute)
Department of Pharmaceutical & Pharmacological Sciences - miscellaneous
× corresponding author
# (joint) last author

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