Ple106-EGFP (Negative)

Ple106-EGFP Description:

The Pleiades Promoter Project created a 1591 bp MiniPromoter derived from two intronic candidate regulatory regions and a segment of the HAP1 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 pEMS1370 with Ple106 driving EGFP. The Ple106-EGFP (pEMS1370) 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. Three ESC lines (mEMS1850, mEMS1852, mESM1858) were PCR validated. Chimeras were generated using mEMS1850.

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

Ple106-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).

Ple106 MiniPromoter Resources:

Ple106-EGFP
	plasmid pEMS1370 (in backbone pEMS1306)
	ESCs mEMS1850, mEMS1852, mEMS1858
	mice generated using ESC mEMS1850
Resource Files
	Ple106 pEMS1370 Sequence	Jones lab
	Ple106 pEMS1370 MiniPromoter Design	Wasserman lab
	Ple106 pEMS1370 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple106 pEMS1370 Vector Map	Holt lab
	Ple106 pEMS1370 Genotyping Assay  (Requires VectorNTI Sofware)	Holt lab

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