@article{Muona2010, abstract = {Segmental isotopic labeling is a powerful labeling technique for reducing nuclear magnetic resonance (NMR) signal overlap, which is associated with larger proteins by incorporating stable isotopes into only one region of a protein for NMR detections. Segmental isotopic labeling can not only reduce complexities of NMR spectra but also retain possibilities to carry out sequential resonance assignments by triple-resonance NMR experiments. We described in vivo (i.e., in Escherichia coli) and in vitro protocols for segmental isotopic labeling of multi-domain and fusion proteins via protein trans-splicing (PTS) using split DnaE intein without any refolding steps or alpha-thioester modification. The advantage of PTS approach is that it can be carried out in vivo by time-delayed dual-expression system with two controllable promoters. A segmentally isotope-labeled protein can be expressed in Escherichia coli within 1 d once required vectors are constructed. The total preparation time of a segmentally labeled sample can be as short as 7-13 d depending on the protocol used.}, author = {Muona, Mikko and Sesilja Aranko, A. and Aranko, A. Sesilja and Raulinaitis, Vytas and Iwaï, Hideo}, doi = {10.1038/nprot.2009.240}, journal = {Nature Protocols}, month = {mar}, pages = {574-587}, title = {Segmental isotopic labeling of multi-domain and fusion proteins by protein trans-splicing in vivo and in vitro}, url = {https://www.researchgate.net/profile/Hideo_Iwai/publication/41761158_Segmental_isotopic_labeling_of_multi-domain_and_fusion_proteins_by_protein_trans-splicing_in_vivo_and_in_vitro/links/02e7e52394a0af3568000000.pdf}, volume = {5}, year = {2010} }