Ple47-EGFP (Negative)

Ple47-EGFP Description:

The Ple47 promoter construct is derived from the promoter region of the CX3 chemokine receptor 1 (CX3CR1)gene. The Pleiades Promoter Project constructed a 3672 bp MiniPromoter from two downstream candidate regulatory regions and a subsection of the CX3CR1 promoter and introduced them into the multiple cloning site (MCS) of Pleiades expression vector pEMS1307 (EGFP reporter flanked by full frt sites) to make plasmid pEMS1152 with Ple47 driving EGFP. The Ple47-EGFP (pEMS1152) 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. Four ESC lines (mEMS3037, mEMS3039, mEMS3048, and mEMS3051) were PCR validated. Chimeras were generated using mEMS3037.

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

Ple47-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).

Ple47 MiniPromoter Resources:

Ple47-EGFP
	plasmid pEMS1152 (in backbone pEMS1307)
	ESCs mEMS3037, mEMS3039, mEMS3048, mEMS3051
	mice generated using ESC mEMS3037
Resource Files
	Ple47 pEMS1152 Sequence	Jones lab
	Ple47 pEMS1152 MiniPromoter Design	Wasserman lab
	Ple47 pEMS1152 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple47 pEMS1152 Vector Map	Holt lab
	Ple47 pEMS1152 Genotyping Assay  (Requires VectorNTI Sofware)	Holt lab

Publications: