Ple148-EGFP (Negative)

Ple148-EGFP Description:

The Ple148 promoter construct is derived from the promoter region of the oligodendrocyte transcription factor 1 (OLIG1) gene. The Pleiades Promoter Project created a 1039 bp MiniPromoter derived from an upstream putative regulatory element and a segment of the promoter, and introduced them into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1169 with Ple148 driving EGFP. The Ple148-EGFP (pEMS1169) 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) mEMS1204.03A. Four ESC lines (mEMS1606, mEMS1611, mEMS1612, and mEMS1624) were PCR validated. Chimeras were generated using mEMS1606.

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

Ple148-EGFP Expression Pattern:

MiniPromoter Ple148 was assayed for EGFP from pEMS1306 backbone. 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).

Ple148 MiniPromoter Resources:

Ple148-EGFP
	plasmid pEMS1169 (in backbone pEMS1306)
	ESCs mEMS1606, mEMS1611, mEMS1612, mEMS1624
	mice generated using ESC mEMS1606
Resource Files
	Ple148 pEMS1169 Sequence	Jones lab
	Ple148 pEMS1169 MiniPromoter Design	Wasserman lab
	Ple148 pEMS1169 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple148 pEMS1169 Vector Map	Holt lab
	Ple148 pEMS1169 Genotyping Assay	Simpson lab
	Ple148 pEMS1169 ESC description	Simpson lab
	Ple148 pEMS1169 Mouse description	Simpson lab

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