The Ple186-EGFP transgene (pEMS1385) was designed with the 3980 bp Ple186 minipromoter (S100b-D;derived from a subsection of the promoter and a downstream putative regulatory element from the human S100 calcium binding protein B (S100B) gene) upstream of a minimal F5 mutant-frt site, an enhanced green fluorescent protein (with mutated TAA->TTA stop), a nuclear localization signal, a second minimal frt site, an SV40 early polyA signal, and a human HPRT complementary sequence (containing exon1, intron1, exon2, and part of intron2). This construct was targeted as a single copy knockin to the Hprtsup>b-m3 mutant locus on the X chromosome.
The promoter/regulatory regions of the human S100 calcium binding protein B (S100B) gene directs expression of enhanced green fluorescent protein (EGFP) to Bergmann glial cells of the cerebellum. Purkinje cells show high background staining. No EGFP signal above background level could be observed in other regions of the brain, such as cortex and brainstem.