URL: http://www.addgene.org/41732
Proper Citation: RRID:Addgene_41732
Insert Name: GltI based Glutamate Biosensor
Organism: Synthetic
Bacterial Resistance: Ampicillin
Defining Citation: PMID:23314171
Vector Backbone Description: Backbone Marker:modified from Invitrogen; Backbone Size:5300; Vector Backbone:pCMV(MinDis) [variant of pDisplay]; Vector Types:Mammalian Expression, Other, Glutamate Biosensor; Bacterial Resistance:Ampicillin
Comments: ** A newer version of this sensor is available. Please see iGluSnFR3 plasmids at https://www.addgene.org/browse/article/28220233/ ** Biosensor iGluSnFRa is a single-wavelength glutamate sensor constructed from E. coli GltI and cpGFP. iGluSnFR is bright and photostable, with 4.5 (ΔF/F)max in vitro, under both 1- and 2-photon illumination. pCMV(MinDis).iGluSnFR was constructed by subcloning the GltI253-cpGFP.L1LV/L2NP gene from pRSET.GltI253-cpGFP.L1LV/L2NP (Addgene plasmid #41733) into the pDisplay vector (Invitrogen) between the BglII and PstI sites. The hemagglutinin (HA) tag was removed by PCR, generating the backbone variant dubbed pCMV(MinDis). This construct encodes an N-terminal mouse immunoglobin κ-chain leader sequence, which directs the protein to the secretory pathway; the GltI253.L1LV/L2NP protein; a myc epitope tag; and at the C-terminus, a platelet derived growth factor receptor transmembrane helix, which anchors the protein to the plasma membrane, displaying it on the extracellular side. Detailed amino acid sequences are provided in the supplementary documents available above. Alternate plasmid names: YbeJ253.L1LVL2NP.pMinDis; pMinDis.iGluSnFR
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Source: Addgene