Tg(Thy1-hop/EYFP)4Gfng

A fusion protein was created with the halophilic bacterium Natronobacterium pharaonisfunctions halorhodopsin (NpHR) fused in-frame with an enhanced yellow fluorescent protein (EYFP). The fusion protein was optimized for expression in mammalian systems and enhanced membrane translocation/reduced bleb formation via C-terminal addition of an endoplasmic reticulum exporting sequence (amino acids 373 to 385 from the potassium channel Kir2.1 gene [NANSFCYENEVALTSK]). This enhanced fusion protein (eNpHR::YFP) was placed downstream of the modified regulatory region of the "murine thy1.2 gene" (extending from the promoter to the intron following exon 4, excluding exon 3 and its flanking introns), and followed by a bovine growth hormone polyA signal. Mice derived from founder line 4 exhibit high EGFP expression in layer 2/3 and layer 5 pyramidal neurons at cortex, dentate gyrus, thalamus, superior colliculus, inferior culliculus, brainstem, amydagala and cerebellum. (J:160662)