Zettler, J., Eppmann, S., Busche, A., Dikovskaya, D., Dötsch, V., Mootz, H.D. and Sonntag, T. (2013) SPLICEFINDER – a fast and easy screening method for active protein trans-splicing positions. PLoS ONE, 8(9), e72925. (doi: 10.1371/journal.pone.0072925) (PMID:24023792) (PMCID:PMC3759424)
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Abstract
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.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Dikovskaya, Dr Dina |
| Authors: | Zettler, J., Eppmann, S., Busche, A., Dikovskaya, D., Dötsch, V., Mootz, H.D., and Sonntag, T. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Cancer Sciences |
| Journal Name: | PLoS ONE |
| Publisher: | Public Library of Science |
| ISSN: | 1932-6203 |
| ISSN (Online): | 1932-6203 |
| Copyright Holders: | Copyright © 2013 The Authors |
| First Published: | First published in PLoS ONE 8(9):e72925 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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