This bacterial two-hybrid system is based on the functional complementation between two complementary fragments, T25 and T18, of the catalytic domain of CyaA, in an E. coli cya strain.
In this study, the effect of cytoplasmic modifications of the A3AR on βarr2 recruitment was evaluated in transiently transfected HEK293T cells, using a live-cell split-reporter system (NanoBit®, Promega), based on the structural complementation of NanoLuc luciferase, allowing real-time βarr2 monitoring.
Following this rational, we recently developed the "split-Cre system" based on the functional complementation of Cre protein fragments [15].
DiCre is based on the functional complementation, using ligand-induced dimerization [25], of two inactive Cre moieties that can be associated by rapamycin through cross-linking the protein fragments FKBP12 (FK506-binding protein; [26]) and FRB, the ligand binding domain of FRAP (FKBP12-rapamycin associated protein; [27]) linked to the Cre moieties.
Split-TEV is based on the functional complementation of two inactive TEV protease fragments fused to interacting proteins.
It is based on the molecular complementation of any enhancer/promoter fragment in the targeting vector with the EGFP reporter gene in the landing site.
To overcome this difficulty, we have developed an in vivo assay based on the transgenic complementation of lat-1 ok1465), lat-1 ok1465tally lethal andevelopmentallyt-1 mutant strain.
Introducing the CDS lengths as a parameter was a good way to simulate the GPI-anchor pathway-mutant screening that was based on the functional complementation assay using candidate gene-expression plasmids.
Importantly, we revealed that KCBP exhibits non-processive movement by the live-cell, single-molecule (particle) motility assay, and that the second MT-binding domain is essential, based on the genetic complementation tests; our data further support the bundling or crosslinking role of KCBP in the formation of the MT rings at the branching site.
