Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System

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15.9K Views

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11:47 min

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August 1st, 2016

DOI :

10.3791/54273-v

August 1st, 2016

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Emanuel G. Worst¹, Matthias P. Exner², Alessandro De Simone², Marc Schenkelberger¹, Vincent Noireaux³, Nediljko Budisa², Albrecht Ott¹

¹Department of Experimental Physics, Saarland University, ²Institute of Chemistry, Technische Universität Berlin, ³School of Physics and Astronomy, University of Minnesota

Podsumowanie

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Keywords Noncanonical Amino Acids

Cell free Expression

Protein Structure

Residue specific Incorporation

Amino Acid Master Mix

CAA

NcAA

Cell free Transcription translation System

Rozdziały w tym wideo

0:05

Title

0:41

Amino Acid Solution Preparation

3:23

Cell-free Reactions for Residue-specific Incorporation of ncAAs

5:06

SDS-PAGE of Cell-free Reactions

6:57

His-tagged Model Protein Purification from the Cell-free Reactions

8:48

Buffer Exchange for HPLC-ESI Mass Spectroscopy

10:03

Results: Successful Incorporation of ncAA in Target Protein

11:14

Conclusion

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