Poliovirus Requires a Precise 5′ End for Efficient Positive-Strand RNA Synthesis
2000; American Society for Microbiology; Volume: 74; Issue: 14 Linguagem: Inglês
10.1128/jvi.74.14.6394-6400.2000
ISSN1098-5514
Autores Tópico(s)Viral Infections and Immunology Research
ResumoABSTRACT Poliovirus infectious RNA can be synthesized in vitro using phage DNA-dependent RNA-polymerases. These synthetic transcripts contain several extra nucleotides at the 5′ end, which are deleted during replication to generate authentic viral genomes. We removed those 5′-end extra nucleotides utilizing a hammerhead ribozyme to produce transcripts with accurate 5′ ends. These transcripts replicate substantially more rapidly in cell culture, demonstrating no lag before replication; they also replicate more efficiently in Xenopus laevis oocytes and in in vitro translation-replication cell extracts. In both systems, an exact 5′ end is necessary for synthesis of positive-strand RNA but not negative-strand RNA.
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