title: A Cassette Approach for the Identification of Intein Insertion Sites.
abstract: Over the past decade split inteins have established themselves as powerful tools for protein engineering, protein semisynthesis, and protein functional control approaches. Their key advantage lies in the protein trans-splicing (PTS) reaction that enables posttranslational protein assembly from two independent, even synthetic, peptide precursors. However, since most split intein applications deal with fragmentation and modification of proteins, various issues can arise, ranging from reduced stability to impairment of protein folding. In this chapter I address how the usage of DNA encoded intein cassettes can streamline and speed up the identification of functional split intein insertion sites in novel proteins of interest (POI).
authors.affiliation: Clayton Foundation Laboratories for Peptide Biology, The Salk Institute for Biological Studies, La Jolla, California, 92037, USA. tsonntag@salk.edu.
authors.forename: Tim
authors.initials: T
authors.lastname: Sonntag
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D047668:Inteins; D015202:Protein Engineering / Q000379:methods*; D019154:Protein Splicing; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins
keywords: CREB; Conditional protein splicing; Homologous recombination; Protein expression; Protein semisynthesis; Protein splicing; RF cloning; Split intein; TEV protease