Journal of Molecular Biology
Product analysis of RNA generated de novo by Qβ replicase
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Applications of phage-derived RNA-based technologies in synthetic biology
2020, Synthetic and Systems BiotechnologyCitation Excerpt :As the simplest (+)-RNA virus, Qβ phage can play a major role in providing the catalytic enzymatic basis for RNA-dependent RNA replication in synthetic systems. Many important results related to Qβ replicase, including the kinetics [105–108], structure [109–111], related host proteins [112–115] and mechanism of template recognition and initiation [116–119], have established Qβ phage as the pioneering model for applications of RNA replication and laid a solid foundation for exploring and employing the RNA replication process. Compared to replicases from other viruses, Qβ replicase has advantages in terms of purification efficiency, activity preservation in cell-free extracts, RNA amplification rate and template specificity [120], which makes Qβ replicase the prime choice for RNA replication in different scenarios.
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2008, Progress in Nucleic Acid Research and Molecular BiologyCitation Excerpt :Yet, after the authors managed to reduce the incubation volume 10,000-fold (down to 20 nl), they still continued to observe spontaneously synthesized RNAs in each individual sample (33). This result gave rise to a hypothesis that efficient Qβ replicase templates are generated de novo by virtue of random nucleotide condensation followed by evolution of fortuitously formed replicable polyribonucleotides into rapidly amplifiable RNA species (34, 35). This hypothesis dominated until 1991, when Qβ replicase version of MCT allowed us to reveal the source of spontaneously generated templates (1).
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