tailieunhanh - Bioluminescence Recent Advances in Oceanic Measurements and Laboratory Applications Part 6

Tham khảo tài liệu 'bioluminescence recent advances in oceanic measurements and laboratory applications part 6', khoa học tự nhiên, công nghệ sinh học phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | 66 Bioluminescence - Recent Advances in Oceanic Measurements and Laboratory Applications galactosidase ubiquitin dihydrofolate reductase firefly luciferase renilla luciferase Gaussia luciferase etc. have been adapted for split-protein strategies. Advantages of this approach include its ability to monitor the interaction of proteins occurring in their specific cellular compartments utility for drug evaluation low background ability to measure real-time interactions applications in protein array analysis and potential adaptability to study both cells and in living animals. However not every reporter protein can be used for this strategy Ozawa et al. 2001 Paulmurugan et al. 2004 Pelletier et al. 1999 Remy and Michnick 1999 Rossi et al. 1997 Wehrman et al. 2002 Figure 5 . Fig. 5. Scheme of a split-reporter protein complementation system for studying proteinprotein interactions. A . Complementation system to study spontaneous protein-protein interactions. In this strategy N- and C- terminal firefly luciferase reporter protein fragments are attached to two study proteins and are expressed as individual fusion proteins N-FLUC-Protein-X and Protein-Y-C-FLUC . When cells are transfected to co-express these two fusion proteins the interaction between protein-X and protein-Y brings the N-FLUC and C-FLUC fragments to a close proximity and induces the complementation to produce reporter signal. B . Complementation system to study small molecule mediated protein-protein interactions. In this strategy N- and C- terminal Renilla luciferase reporter protein fragments are attached to two study proteins and are expressed as individual fusion proteins N-hRLUC-Protein-A and Protein-B-C-hRLUC . When cells are transfected to coexpress these two fusion proteins the interaction between protein-A and protein-B is induced by a small molecule drug which brings the N-hRLUC and C-hRLUC fragments to a close proximity and causes the complementation to produce reporter signal. In both strategies .