title: SPLICEFINDER - a fast and easy screening method for active protein trans-splicing positions.
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.
authors.affiliation: Department of Chemistry and Chemical Biology, TU Dortmund University, Dortmund, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Zettler
authors.forename: Simone
authors.initials: S
authors.lastname: Eppmann
authors.forename: Alena
authors.initials: A
authors.lastname: Busche
authors.forename: Dina
authors.initials: D
authors.lastname: Dikovskaya
authors.forename: Volker
authors.initials: V
authors.lastname: Dötsch
authors.forename: Henning D
authors.initials: HD
authors.lastname: Mootz
authors.forename: Tim
authors.initials: T
authors.lastname: Sonntag
mesh_terms: D019154:Protein Splicing / Q000235:genetics*; D011506:Proteins / Q000737:chemistry* / Q000235:genetics*; D020040:Trans-Splicing / Q000235:genetics*
chemical_list: D011506:Proteins
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