N-Terminal Amino Acid Affects the Translation Efficiency at Lower Temperatures in a Reconstituted Protein Synthesis System
Abstract
:1. Introduction
2. Results
2.1. Synthesis of Model Proteins at Different Temperatures
2.2. Synthesis of sfGFP with a Different Fourth Amino Acid
2.3. Effect of a Synonymous Codon at the Fourth Position
2.4. Synthesis of sfGFP with Different Amino Acids at the Second Position
2.5. Effect of Addition of N-Terminal Sequence on the Productivity of Other Proteins
2.6. Expression of sfGFP Variants in E. coli
3. Discussion
4. Materials and Methods
4.1. Preparation of Template and Plasmid DNA for Cell-Free Protein Synthesis and Expression in E. coli, Respectively
4.2. Protein Synthesis Using the PURE System
4.3. Expression of sfGFP in E. coli
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Fuse-Murakami, T.; Matsumoto, R.; Kanamori, T. N-Terminal Amino Acid Affects the Translation Efficiency at Lower Temperatures in a Reconstituted Protein Synthesis System. Int. J. Mol. Sci. 2024, 25, 5264. https://doi.org/10.3390/ijms25105264
Fuse-Murakami T, Matsumoto R, Kanamori T. N-Terminal Amino Acid Affects the Translation Efficiency at Lower Temperatures in a Reconstituted Protein Synthesis System. International Journal of Molecular Sciences. 2024; 25(10):5264. https://doi.org/10.3390/ijms25105264
Chicago/Turabian StyleFuse-Murakami, Tomoe, Rena Matsumoto, and Takashi Kanamori. 2024. "N-Terminal Amino Acid Affects the Translation Efficiency at Lower Temperatures in a Reconstituted Protein Synthesis System" International Journal of Molecular Sciences 25, no. 10: 5264. https://doi.org/10.3390/ijms25105264