ArticleSynthetic and systems biotechnology2021
A linear DNA template-based framework for site-specific unnatural amino acid incorporation.
Article in Synthetic and systems biotechnology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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3 citing papers in PubMed.
- Customized synthesis of phosphoprotein bearing phosphoserine or its nonhydrolyzable analog.Synthetic and systems biotechnology · 2023Article
- Toward efficient multiple-site incorporation of unnatural amino acids using cell-free translation system.Synthetic and systems biotechnology · 2022Article
- Cell-free synthetic biology: Orchestrating the machinery for biomolecular engineering.Biotechnology notes (Amsterdam, Netherlands) · 2022Article
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4 authors.
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Abstract
Site-specific incorporation of unnatural amino acids (UNAAs) into proteins using an orthogonal translation system (OTS) has expanded the scope of protein-coding chemistry. The key factor affecting UNAA embedding efficiency is the orthogonality of the OTS. Compared to traditional cell systems, cell-free systems are more convenient to control the reaction process and improve the utilization rate of UNAA. In this study, a linear DNA template-based cell-free unnatural protein synthesis system for rapid high-throughput screening and evolution was proposed. A total of 14 cell extracts were selected for screening out cell extract with high expression level. The result showed that EcAR7 ΔA ΔSer cell extract was optimal for the cell-free system. In addition, the screening results of four UNAAs, p-propargyloxy-l-phenylalanine (pPaF), p-azyl-phenylalanine (pAzF), p-acetyl-l-phenylalanine (pAcF), and p-benzoyl-l-phenylalanine (pBpF), showed that o-aaRS and o-tRNA of pPaF had good orthogonality. A new pair of corresponding o-aaRS and o-tRNA for pBpF was screened out. These results proved that this method could speed up the screening of optimal OTS components for UNAAs with versatile functions.
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