Derivation of an infectious viral RNA by autolytic cleavage of in vitro transcribed viral cDNAs.
- 1 July 1989
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 86 (13) , 4823-4827
- https://doi.org/10.1073/pnas.86.13.4823
Abstract
An in vitro transcription system has been developed that utilizes self-processing to adjust 3'' termini in transcribed viral RNAs. Transcription vectors were constructed by ligating a cDNA "cassette" containing self-cleavage sequences from the satellite of tobacco ringspot virus (STobRV RNA) with the 3'' end of brome mosaic virus (BMV) cDNA clones. Transcripts were obtained either from templates linearized at positions located downstream from the cassette or from nonlinearized plasmids. In both cases, a spontaneous self-cleavage reaction produced BMV RNAs that contained 19 heterologous nucleotides at their 3'' ends. These RNAs were biologically active, indicating that 19 extra nucleotides did not remove the infectivity. The progeny BMV RNA isolated from infected plants did not retain the 19-base heterologous sequence. BMV RNAs containing longer 3'' heterologous sequences were not infectious. The STobRV self-cleavage cassette is potentially applicable to a variety of RNA systems.Keywords
This publication has 26 references indexed in Scilit:
- [19] Rapid and efficient site-specific mutagenesis without phenotypic selectionPublished by Elsevier ,2004
- BIOLOGICAL CATALYSIS BY RNAAnnual Review of Biochemistry, 1986
- Infectious RNA derived by transcription from cloned cDNA copies of the genomic RNA of an insect virus.Proceedings of the National Academy of Sciences, 1986
- Molecular cloning of the genome of a cardiotropic Coxsackie B3 virus: full-length reverse-transcribed recombinant cDNA generates infectious virus in mammalian cells.Proceedings of the National Academy of Sciences, 1985
- Mutant viral RNAs synthesized in vitro show altered aminoacylation and replicase template activitiesNature, 1984
- Efficientin vitrosynthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoterNucleic Acids Research, 1984
- Organization and Expression of Bacteriophage T7 DNACold Spring Harbor Symposia on Quantitative Biology, 1983
- Bacteriophage SP6-specific RNA polymerase. I. Isolation and characterization of the enzyme.Journal of Biological Chemistry, 1982
- †RNA-like Structures in the Genomes of RNA VirusesProgress in Nucleic Acid Research and Molecular Biology, 1982
- Cloned Poliovirus Complementary DNA Is Infectious in Mammalian CellsScience, 1981