DNA Template Preparation by PCR for In Vitro Transcription
3:20
mRNA Generation by In Vitro Transcription
4:22
Cells Transfection with mRNA
6:04
Luciferase Activity Measurement
7:29
Results: Translation Efficiency of a Fluc mRNA Determined by Luciferase Reporter Assay
8:21
Conclusion
TRANSCRIÇÃO
Our protocol for an in vitro transcribed RNA-based luciferase assay helps study translation regulation in pox virus infected cells. Additionally, a customized length of the Poly(A)liter allows studying its regulatory effects on translation. The ma
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We present a protocol to study mRNA translation regulation in poxvirus-infected cells using in vitro Transcribed RNA-based luciferase reporter assay. The assay can be used for studying translation regulation by cis-elements of an mRNA, including 5’-untranslated region (UTR) and 3’-UTR. Different translation initiation modes can also be examined using this method.