Journal
VIROLOGY
Volume 446, Issue 1-2, Pages 230-237Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2013.07.038
Keywords
Filovirus; Marburgvirus; Marburg virus; Bat isolate; Minigenome; Recombinant; Full-length; GFP; Macrophages; Cytokines
Categories
Funding
- Centers for Disease Control and Prevention (CDC)
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Recent investigations have shown the Egyptian fruit bat (Rousettus aegyptiacus) to be a natural reservoir for marburgviruses. To better understand the life cycle of these viruses in the natural host, a new reverse genetics system was developed for the reliable rescue of a Marburg virus (MARV) originally isolated directly from a R. aegyptiacus bat (371 Bat). To develop this system, the exact terminal sequences were first determined by 5' and 3' RACE, followed by the cloning of viral proteins NP, VP35, VP30 and L into expression plasmids. Novel conditions were then developed to efficiently replicate virus mini-genomes followed by the construction of full-length genomic clones from which recombinant wild type and GFP-containing MARVs were rescued. Surprisingly, when these recombinant MARVs were propagated in primary human macrophages, a dramatic difference was found in their ability to grow and to elicit antiviral cytokine responses. Published by Elsevier Inc.
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