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体外转录 rna 基荧光素酶记者法研究类病毒感染细胞的翻译规律
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
In Vitro Transcribed RNA-based Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-infected Cells
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体外转录 rna 基荧光素酶记者法研究类病毒感染细胞的翻译规律

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

Capitoli

  • 00:04Titolo
  • 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

Traduzione automatica

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我们提出了一种研究 mrna 翻译调节的方案, 在天花病毒感染细胞使用体外转录 Rna 基荧光素酶报告法。该方法可用于研究 mRNA 的cis元素的翻译调控, 包括 5 '-未翻译区域 (utr) 和 3 '-utr。使用此方法还可以检查不同的翻译启动模式。

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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