Genetic Code Expansion, Protein Expression, and Protein Functionalization in Bacillus subtilis
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
- MeSH
- Bacillus subtilis genetika metabolismus MeSH
- click chemie MeSH
- CRISPR-Cas systémy MeSH
- ELISA MeSH
- genetické vektory MeSH
- genetický kód MeSH
- isopropylthiogalaktosid farmakologie MeSH
- kreatinkinasa, forma MM metabolismus MeSH
- lidé MeSH
- lysin chemie MeSH
- norbornany chemie MeSH
- proteinové inženýrství metody MeSH
- regulace genové exprese u bakterií účinky léků MeSH
- rekombinantní proteiny chemie genetika izolace a purifikace metabolismus MeSH
- zelené fluorescenční proteiny genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 2-norbornene MeSH Prohlížeč
- isopropylthiogalaktosid MeSH
- kreatinkinasa, forma MM MeSH
- lysin MeSH
- norbornany MeSH
- rekombinantní proteiny MeSH
- zelené fluorescenční proteiny MeSH
The site-specific chemical modification of proteins through incorporation of noncanonical amino acids enables diverse applications, such as imaging, probing, and expanding protein functions, as well as to precisely engineer therapeutics. Here we report a general strategy that allows the incorporation of noncanonical amino acids into target proteins using the amber suppression method and their efficient secretion in the biotechnological relevant expression host Bacillus subtilis. This facilitates efficient purification of target proteins directly from the supernatant, followed by their functionalization using click chemistry. We used this strategy to site-specifically introduce norbornene lysine into a single chain antibody and functionalize it with fluorophores for the detection of human target proteins.
Citace poskytuje Crossref.org
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