Hprt1^{tm32(Ple112-EGFP)Ems}

The Ple112-EGFP transgene (pEMS1418) was designed with the 1295 bp Ple112 minipromoter (HCRT-B; derived from a subsection of the promoter and an upstream putative regulatory element from the human hypocretin (orexin) neuropeptide precursor (HCRT) 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 Hprtb-m3 mutant locus on the X chromosome. The promoter/regulatory regions of the human hypocretin (HCRT) gene directs expression of enhanced green fluorescent protein (EGFP) to the hypothalamus. Under the higher magnification, EGFP immunoreactivity can be detected in the cell bodies and processes of hypothalamic neurons. Other (lateral) regions of brain did not display any EGFP-positive cells. (J:144244, J:164356)