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- W2935880354 abstract "Abstract Protein‐splicing domains are frequently used engineering tools that find application in the in vivo and in vitro ligation of protein domains. Directed evolution is among the most promising technologies used to advance this technology. However, the available screening systems for protein‐splicing activity are associated with bottlenecks such as the selection of pseudo‐positive clones arising from off‐pathway reaction products or fragment complementation. Herein, we report a stringent screening method for protein‐splicing activity in cis and trans , that exclusively selects productively splicing domains. By fusing splicing domains to an intrinsically disordered region of the antidote from the Escherichia coli CcdA/CcdB type II toxin/antitoxin system, we linked protein splicing to cell survival. The screen allows selecting novel cis ‐ and trans ‐splicing inteins catalyzing productive highly efficient protein splicing, for example, from directed‐evolution approaches or the natural intein sequence space." @default.
- W2935880354 created "2019-04-25" @default.
- W2935880354 creator A5026837905 @default.
- W2935880354 creator A5070551735 @default.
- W2935880354 date "2019-07-03" @default.
- W2935880354 modified "2023-10-07" @default.
- W2935880354 title "Off‐Pathway‐Sensitive Protein‐Splicing Screening Based on a Toxin/Antitoxin System" @default.
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- W2935880354 doi "https://doi.org/10.1002/cbic.201900139" @default.
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