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In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme
DOI:

12:36 min

October 19, 2010

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Chapters

  • 00:00Title
  • 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

Automatic Translation

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.

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