Ple45-EGFP (Negative)

Ple45-EGFP Description:

The Ple45 promoter construct is derived from the promoter region of the CX3 chemokine receptor 1 (CX3CR1) gene. The Pleiades Promoter Project constructed a 1620 bp MiniPromoter from two candidate regulatory regions of the CX3CR1 gene along with 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 pEMS1150 with Ple45 driving EGFP. The Ple45-EGFP (pEMS1150) 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 (mEMS3035, mEMS3071, mEMS3072, and mEMS3092) were PCR validated. Chimeras were generated using mEMS3035.

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

Ple45-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).

Ple45 MiniPromoter Resources:

Ple45-EGFP
	plasmid pEMS1150 (in backbone pEMS1307)
	ESCs mEMS3035, mEMS3071, mEMS3072, mEMS3092
	mice generated using ESC mEMS3035
Resource Files
	Ple45 pEMS1150 Sequence	Jones lab
	Ple45 pEMS1150 MiniPromoter Design	Wasserman lab
	Ple45 pEMS1150 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple45 pEMS1150 Vector Map	Holt lab
	Ple45 pEMS1150 Genotyping Assay  (Requires VectorNTI Sofware)	Holt lab

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