Figure 6
From: A New Protein-Protein Interaction Sensor Based on Tripartite Split-GFP Association
Monitoring complex formation and stability in E. coli.
(a) Analysis of YheNML heterotrimeric complex formation using split-GFP. GFP1–9 complementation provides information on interaction of GFP10 and GFP11 tagged domains, while GFP1–10 binds to GFP11 tagged protein, thus providing expression and solubility levels of the GFP11 tagged protein or complex. (b) E. coli polycistrons YheNML, YheML and YBJG/K were subcloned between GFP10 and GFP11 strands with extended linkers (l25 and l30) into pTET GFP10/11 tet inducible plasmids. (c) Split-GFP assay in E. coli cells expressing either GFP1–9 or GFP1–10. Sequential or co-induction was performed as previously described. Corresponding semi-quantitative measure of colony fluorescence using NIH Image (right).
