Infectious RNA Transcripts from Ross River Virus cDNA Clones and the Construction and Characterization of Defined Chimeras with Sindbis Virus
Abstract
We have constructed a full-length cDNA clone of the virulent T48 strain of Ross River virus, a member of the alphavirus genus. Infectious RNA can be transcribed from this clone using SP6 or T7 RNA polymerase. The rescued virus has properties indistinguishable from those of the T48 strain of Ross River virus. We have used this clone, together with a full-length cDNA clone of Sindbis virus, to construct chimeric plasmids in which the 5' and the 3' nontranslated regions of the Sindbis and Ross River genomes were exchanged. The nontranslated regions of the two viral genomes differ in both size and sequence although they maintain specific conserved sequence elements. Virus was recovered from all four chimeras. Chimeras containing heterologous 3' nontranslated regions had replicative efficiencies equal to those of the parents. In contrast, the chimeras containing heterologous 5' nontranslated regions were defective in RNA synthesis and virus production, and the severity of the defect was dependent upon the host. Replication of a virus containing a heterologous 5' nontranslated region may be inefficient due to the formation of defective protein-RNA complexes, whereas, the presumptive complexes formed between host or virus proteins and the 3' nontranslated region to promote RNA synthesis appear to function normally in the chimeras.
Additional Information
© 1991 Academic Press, Inc. Received December 20, 1990; accepted February 12, 1991. We thank Frank Preugschat and Ellen Strauss for stimulating discussions. We also thank Ellen Strauss for critical reading of the manuscript. This work was supported by Grant DMB-8617372 from the National Science Foundation and by Grants AI 10793 and AI 20712 from the National Institutes of Health. R.J.K. was a recipient of a U.S. Public Health Service Individual National Research Service Award (AI07869) from the National Institutes of Health. H.G.M.N. was supported in part by a fellowship grant from the Niels Stensen Foundation, a Gosney Fellowship from CIT, and a travel grant from The Netherlands Organization for the Advancement of Pure Research.Additional details
- Eprint ID
- 64660
- DOI
- 10.1016/0042-6822(91)90584-X
- Resolver ID
- CaltechAUTHORS:20160222-154455299
- DMB-8617372
- NSF
- Al 10793
- NIH
- Al 20712
- NIH
- AI 07869
- NIH
- Niels Stensen Foundation (NSS)
- Caltech Gosney Fellowship
- Nederlandse Organisatie voor Zuiver-Wetenschappelijk Onderzoek (ZWO)
- Created
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2016-02-23Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field