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Остаток конкретных Включение неканонических аминокислот в модельных белков с использованием<em> Кишечная палочка</em> Бесклеточной системы транскрипции-трансляции
JoVE Journal
Biology
This content is Free Access.
JoVE Journal Biology
Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System
DOI:

11:47 min

August 01, 2016

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Chapters

  • 00:05Title
  • 00:41Amino Acid Solution Preparation
  • 03:23Cell-free Reactions for Residue-specific Incorporation of ncAAs
  • 05:06SDS-PAGE of Cell-free Reactions
  • 06:57His-tagged Model Protein Purification from the Cell-free Reactions
  • 08:48Buffer Exchange for HPLC-ESI Mass Spectroscopy
  • 10:03Results: Successful Incorporation of ncAA in Target Protein
  • 11:14Conclusion

Summary

Automatic Translation

An easy-to-use, cell-free expression protocol for the residue-specific incorporation of noncanonical amino acid analogs into proteins, including downstream analysis, is presented for medical, pharmaceutic, structural and functional studies.

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