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Immunology and Infection
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In Vitro Transcribed RNA-based Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-infected Cells
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Immunology and Infection
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JoVE Journal Immunology and Infection
In Vitro Transcribed RNA-based Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-infected Cells

In Vitro Transcribed RNA-based Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-infected Cells

DOI:
10.3791/59626-v

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08:58 min

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May 01, 2019

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, , ,
1Division of Biology,Kansas State University

Chapters

  • 00:04Title
  • 00:42DNA Template Preparation by PCR for In Vitro Transcription
  • 03:20mRNA Generation by In Vitro Transcription
  • 04:22Cells Transfection with mRNA
  • 06:04Luciferase Activity Measurement
  • 07:29Results: Translation Efficiency of a Fluc mRNA Determined by Luciferase Reporter Assay
  • 08:21Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

In Vitro Transcribed RNALuciferase Reporter AssayTranslation RegulationPoxvirus-infected CellsPoly(A) Tail5’UTR3’UTRCap ModificationPCR AmpliconT7 PromoterFirefly LuciferaseRenilla LuciferaseKozak SequenceAgarose Gel Electrophoresis

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