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In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme
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Immunologie et infection
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Journal JoVE Immunologie et infection
In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme

In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme

DOI:
10.3791/2390-v

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12:36 min

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October 19, 2010

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, , ,
1Faculty of Medicine, Rappaport Institute,Technion – Israel Institute of Technology, 2The Pharmacy and Biochemistry Institute,Johannes Gutenberg University

Chapitres

  • 00:00Titre
  • 01:43tRNA Preparation
  • 05:28Expression and Purification of the Recombinant E. histolytica Dnmt2 (Ehmeth) Protein in E. coli BL21
  • 08:06 in vitro tRNA Methylation Assay
  • 10:30Representative Results
  • 12:02Conclusion

Summary

Traduction automatique

Traduction automatique

This protocol describes the preparation of a synthetic tRNA substrate for the Entamoeba histolytica DNA/tRNA methyltransferase 2 (Dnmt2) homolog Ehmeth and the measure of its methyltransferase activity. This experimental approach can be used for investigating the activity of other Dnmt2 proteins.

Tags

In Vitro TRNA Methylation AssayEntamoeba HistolyticaDNA Methyltransferase Dnmt2Ehmeth EnzymeProtozoan ParasitesAmebiasisInfectious AgentsClinical Manifestations5-methylcytosine (5mC)Single DNA MethyltransferaseControl Of Repetitive ElementsDual DNA/tRNAAsp Methyltransferase ActivityFunctional SignificanceTRNA Methyltransferase ActivityDnmt2 Proteins

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