Ontology highlight
ABSTRACT:
SUBMITTER: Dhungel P
PROVIDER: S-EPMC7236376 | biostudies-literature | 2019 May
REPOSITORIES: biostudies-literature
Journal of visualized experiments : JoVE 20190501 147
Every poxvirus mRNA transcribed after viral DNA replication has an evolutionarily conserved, non-templated 5'-poly(A) leader in the 5'-UTR. To dissect the role of 5'-poly(A) leader in mRNA translation during poxvirus infection we developed an in vitro transcribed RNA-based luciferase reporter assay. This reporter assay comprises of four core steps: (1) PCR to amplify the DNA template for in vitro transcription; (2) in vitro transcription to generate mRNA using T7 RNA polymerase; (3) Transfection ...[more]