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 acti
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.
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Metadaten
Author:Joachim Zettler, Eppmann Simone, Alena Busche, Dina Dikovskaya, Volker Dötsch, Henning D. Mootz, Tim Sonntag
URN:urn:nbn:de:hebis:30:3-315808
DOI:http://dx.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
Pagenumber: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 und Chemie
Dewey Decimal Classification:570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Licence (German):License LogoCreative Commons - Namensnennung 3.0

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