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- MI_0111 IAO_0000115 "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 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." @default.
- MI_0111 normalizedInformationContent "100" @default.
- MI_0111 referenceCount "1" @default.
- MI_0111 hasExactSynonym "dhfr reconstruction" @default.
- MI_0111 hasOBONamespace "PSI-MI" @default.
- MI_0111 id "MI:0111" @default.
- MI_0111 inSubset PSI-MI_slim @default.
- MI_0111 type Class @default.
- MI_0111 isDefinedBy mi.owl @default.
- MI_0111 label "dihydrofolate reductase reconstruction" @default.
- MI_0111 subClassOf MI_0000 @default.
- MI_0111 subClassOf MI_0001 @default.
- MI_0111 subClassOf MI_0045 @default.
- MI_0111 subClassOf MI_0090 @default.
- MI_0111 subClassOf MI_0111 @default.