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Splicefinder : a fast and easy screening method for active protein trans-splicing positions
- Split intein enabled protein trans-splicing (PTS) is a powerful method for the ligation of two protein fragments, thereby paving the way for various protein modification or protein function control applications. PTS activity is strongly influenced by the amino acids directly flanking the splice junctions. However, to date no reliable prediction can be made whether or not a split intein is active in a particular foreign extein context. Here we describe SPLICEFINDER, a PCR-based method, allowing fast and easy screening for active split intein insertions in any target protein. Furthermore we demonstrate the applicability of SPLICEFINDER for segmental isotopic labeling as well as for the generation of multi-domain and enzymatically active proteins.
| Author: | Joachim Zettler, Eppmann Simone, Alena Busche, Dina Dikovskaya, Volker DötschORCiDGND, Henning D. Mootz, Tim Sonntag |
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| URN: | urn:nbn:de:hebis:30:3-315808 |
| DOI: | https://doi.org/10.1371/journal.pone.0072925 |
| ISSN: | 1932-6203 |
| Parent Title (English): | PLoS One |
| Publisher: | PLoS |
| Place of publication: | Lawrence, Kan. |
| Document Type: | Article |
| Language: | English |
| Date of Publication (online): | 2013/09/02 |
| Date of first Publication: | 2013/09/02 |
| Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
| Release Date: | 2013/09/06 |
| Volume: | 8 |
| Issue: | (9):e72925 |
| Page Number: | 9 |
| Note: | Copyright: © 2013 Zettler et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
| HeBIS-PPN: | 353145645 |
| Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
| Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
| Sammlungen: | Universitätspublikationen |
| Licence (German): |  Creative Commons - Namensnennung 3.0 |
