Flavivirus RNA synthesis in vitro

作者：Padmanabhan Radhakrishnan^*; Takhampunya Ratree; Teramoto Tadahisa; Choi Kyung H

来源：Methods, 2015, 91: 20-34.

DOI：10.1016/j.ymeth.2015.08.002

• RNA-dependent RNA polymerase from positive strand mosquito-borne Flaviviridae
• Cytoplasmic membrane-bound viral replication complex
• Endogenous and exogenous viral RNA templates
• Temperature-dependent de novo initiation and elongation
• Heparin trap
• Polarity of RNA product by RNase H mapping

Establishment of in vitro systems to study mechanisms of RNA synthesis for positive strand RNA viruses have been very useful in the past and have shed light on the composition of protein and RNA components, optimum conditions, the nature of the products formed, cis-acting RNA elements and trans-acting protein factors required for efficient synthesis. In this review, we summarize our current understanding regarding the requirements for flavivirus RNA synthesis in vitro. We describe details of reaction conditions, the specificity of template used by either the multi-component membrane-bound viral replicase complex or by purified, recombinant RNA-dependent RNA polymerase. We also discuss future perspectives to extend the boundaries of our knowledge.

• 出版日期2015-12