Patent ID: 8394604
Filing Date: 2013-03-12
Classification: C07K,C12P

Abstract:
1. A method for covalently linking the N-terminus of a peptide to the C-terminus of a peptide, the method comprising providing a C-terminus of a peptide, the C-terminus being covalently linked via a peptide bond to an N-terminal split intein (In); providing an N-terminus of a peptide, the N-terminus being covalently linked via a peptide bond to a C-terminal split intein (Ic); wherein the Ic is from 4 to 7 contiguous amino acids, at least 4 of which are from the last beta-strand of the C-terminal region of the Ssp GyrB intein, the Ssp DnaX intein, the Ter DnaE3 intein, or the Ter ThyX intein; and wherein the Ic is split from the intein at a site between the penultimate beta-strand and the last beta-strand of the C-terminal region of the intein; and contacting the In-linked C-terminus with the Ic-linked N-terminus in the presence of an intein-splicing polypeptide (ISP) under conditions allowing splicing of the In and the Ic and linking of the C-terminus to the N-terminus; wherein the In, the Ic and the ISP are of the same or cross-reacting intein (I); and wherein the In is from 11 to 20 contiguous amino acids of the N-terminal region of the intein (I); and is split from the intein (I) at a site next to the first or the second beta-strand; wherein the N-terminus and the C-terminus belong to the same peptide, thereby resulting in a cyclic peptide.