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As decide in the PSI genevra meeting MI:0347 is no more obsolete and rename protein tri hybrid. A generic term that is parent of both protein tri hybrid and rna tri hybrid has been created. See MI:0588. Dihydrofolate reductase reconstruction MI:0111 is not considered a 3h method as the 3rd element is a colored substrate of a reconstructed enzyme not a binding partner of the bait and the prey.
Original comment by: luisa_montecchi
3-Hybrid
A technic to show a ternary complex :
PMID : 16080001
It differs from:
term: tri hybrid id: MI:0437 definition: cDNA encoding the bait, the prey and a third protein component is transfected into a recombinant yeast strain for library transformation. \nOBSOLETE this is a two hybrid variation remap to "two hybrid" (MI:0018) , or to "rna tri hybrid" (MI:0438) definition_reference: PMID:12761205 definition_reference: PMID:12935900 definition_reference: PMID:12052864
term: dhfr reconstruction: dihydrofolate reductase reconstruction id: MI:0111 definition: The gene for DHFR is rationally dissected into two fragments called F[1,2] and F[3]. Two proteins or protein domains that are thought to bind to each other can then be fused to either of the two DHFR fragments. Reconstitution of enzyme activity can be monitored in vivo by cell survival in DHFR-negative cells grown in the absence of nucleotides. A fluorescence assay can also be carried out taking advantage of fMTX (the third element of the so called triple hybrid essay) binding to reconstituted DHFR. The basis of this assay is that complementary fragments of DHFR, when expressed and reassembled in cells, will bind with high affinity (Kd 5 540 pM) to fMTX in a 1:1 complex. fMTX is retained in cells by this complex, whereas the unbound fMTX is actively and rapidly transported out of the cells. Survival depends only on the number of molecules of DHFR reassembled. definition_reference: PMID:10318894
term: rna tri hybrid id: MI:0438 definition: In vivo reconstruction of specific RNA-proteins interactions. The DNA binding and transcription activator domains of GAL4 are brought together via the interaction of recombinant RNA. The first hybrid protein contains the DNA binding domain of GAL4 fused to RevM10 (a mutated RNA binding protein of HIV-1 that binds specifically to the Rev responsive element RRE of the env gene). A recombinant RNA contains the RRE sequence and a target RNA sequence X. The second hybrid protein contains the activation domain of GAL4 fused to protein Y tested for its ability to bind the target RNA X. If this interaction occurs the three hybrid
reconstructs GAL4 and the transcription of a reporter gene is activated. definition_reference: PMID:8972875 definition_reference: PMID:12162957
Karine
Reported by: krobbe4