[EN] ONE STEP N-TERMINAL TAGGING OF PROTEINS<br/>[FR] MARQUAGE N-TERMINAL EN UNE SEULE ÉTAPE DES PROTÉINES
申请人:UNIV PENNSYLVANIA
公开号:WO2013188607A1
公开(公告)日:2013-12-19
A selective method of modifying the N-terminus a protein using an aminoacyl tRNA transferase is provided. The method comprises reacting the protein with the aminoacyl adenosyl derivative, recognizable by AaT t-RNA transferase derived from E.Coli and similar bacterial systems. The method further includes production of protein fusion constructs by incorporation of an additional cysteine in the N -terminus and reacting this cysteine with a thioether placed on the C-terminus of a reaction partner protein, producing a fused construct. Selenocysteine can be included in the fused construct and further reduced producing alanine residue in the desired positions. An essential feature of the method is that the aminoacyl adenosine derivatives are pre-made and do not require a synthetase enzyme, allowing a great diversity of the N-terminus modifiers to be pre-made synthetically. The method is implemented as a kit.