| Allele Name | targeted mutation 32, Elizabeth M Simpson |
| Allele Type | Targeted (Reporter) |
| Allele Synonym(s) | HCRT-EGFP; Hprttm32(mEMS1736)Ems; Ple112 |
| Gene Symbol and Name | Hprt, hypoxanthine guanine phosphoribosyl transferase |
| Gene Synonym(s) | |
| Promoter | HCRT, hypocretin neuropeptide precursor, human |
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| Expressed Gene | GFP, Green Fluorescent Protein, |
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| Site of Expression | EGFP is expressed in the hypothalamus. |
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| Strain of Origin | (B6.129P2-Hprtb-m3/J x 129S-Gt(ROSA)26Sortm1Sor/J)F1 |
| Chromosome | X |
| General Note | Germ line transmission of mutant cell line mEMS1736 has been established. |
| Molecular Note | 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. |
| Mutations Made By | Elizabeth Simpson, Centre for Molecular Medicine & Therapeutics, University of British Columbia |
