Document Type
Article
Publication Version
Published Version
Publication Date
2014
Journal or Book Title
Virology Journal
Volume
11
First Page
150-1
Last Page
150-12
DOI
10.1186/1743-422X-11-150
Abstract
Background: Most known flaviviruses, including West Nile virus (WNV), are maintained in natural transmission cycles between hematophagous arthropods and vertebrate hosts. Other flaviviruses such as Modoc virus (MODV) and Culex flavivirus (CxFV) have host ranges restricted to vertebrates and insects, respectively. The genetic elements that modulate the differential host ranges and transmission cycles of these viruses have not been identified. Methods: Fusion polymerase chain reaction (PCR) was used to replace the capsid (C), premembrane (prM) and envelope (E) genes and the prM-E genes of a full-length MODV infectious cDNA clone with the corresponding regions of WNV and CxFV. Fusion products were directly transfected into baby hamster kidney-derived cells that stably express T7 RNA polymerase. At 4 days post-transfection, aliquots of each supernatant were inoculated onto vertebrate (BHK-21 and Vero) and mosquito (C6/36) cells which were then assayed for evidence of viral infection by reverse transcription-PCR, Western blot and plaque assay. Results: Chimeric virus was recovered in cells transfected with the fusion product containing the prM-E genes of WNV. The virus could infect vertebrate but not mosquito cells. The in vitro replication kinetics and yields of the chimeric virus were similar to MODV but the chimeric virus produced larger plaques. Chimeric virus was not recovered in cells transfected with any of the other fusion products. Conclusions: Our data indicate that genetic elements outside of the prM-E gene region of MODV condition its vertebrate-specific phenotype.
Rights
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Copyright Owner
Saiyasombat et al
Copyright Date
2014
Language
en
File Format
application/pdf
Recommended Citation
Saiyasombat, Rungrat; Carrillo-Tripp, Jimena; Miller, W. Allen; Bredenbeek, Peter J.; and Blitvich, Bradley J., "Substitution of the premembrane and envelope protein genes of Modoc virus with the homologous sequences of West Nile virus generates a chimeric virus that replicates in vertebrate but not mosquito cells" (2014). Plant Pathology and Microbiology Publications. 43.
https://lib.dr.iastate.edu/plantpath_pubs/43
Included in
Agricultural Science Commons, Agriculture Commons, Microbiology Commons, Plant Pathology Commons, Veterinary Microbiology and Immunobiology Commons
Comments
This article is from Virology Journal 11 (2014): 150, doi: 10.1186/1743-422X-11-150. Posted with permission.