Ple135-EGFP (Negative)

Ple135-EGFP Description:

The Ple135 promoter construct is derived from the promoter region of the Orphan nuclear receptor NR2E1 (NR2E1) gene. The Pleiades Promoter Project constructed a 1638 bp MiniPromoter from a subsection of the NR2E1 promoter and introduced it into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1351 with Ple135 driving EGFP. The Ple135-EGFP (pEMS1351) was introduced by homologous recombination as a single copy insertion into the Hprt1b-m3 locus in the X chromosome of mouse embryonic stem cells (ESCs) mEMS1204. One ESC line (mMES2100) was PCR validated, and chimeras were generated using it.

Note: Promoterless negative controls (pEMS1302, pEMS1306, pEMS1313) have been generated, as well as CAG positive controls (pEMS1157, pEMS1277 & pEMS1488).

Ple135-EGFP Expression Pattern:

No EGFP expression was detected above background in native fluorescence under epi or confocal microscopy. Anti GFP immunostaining (rabbit polyclonal, Invitrogen) did not detect specific GFP expression above background across numerous brain regions (brightfield).

Ple135 MiniPromoter Resources:

Ple135-EGFP
	plasmid pEMS1351 (in backbone pEMS1306)
	ESCs mEMS2100
	mice generated using ESC mEMS2100
Resource Files
	Ple135 pEMS1351 Sequence	Jones lab
	Ple135 pEMS1351 MiniPromoter Design	Wasserman lab
	Ple135 pEMS1351 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple135 pEMS1351 Vector Map	Holt lab

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