VV064: 1xCox8 - ratiometric pHluorin in fck
PurposeExpresses the pH sensor ratiometric pHluorin in the mitochondria of mammalian cells
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||58502||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backboneFCK(1.3)GW, from Addgene 22217
- Backbone size w/o insert (bp) 9240
- Total vector size (bp) 10059
Vector typeMammalian Expression, Lentiviral
Growth in Bacteria
Growth instructionsUse recombinase-free E. coli.
Gene/Insert name1xCox8 - Ratiometric pHluorin
Insert Size (bp)820
- Promoter a-CamKII
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer GCTCGTCAATCAAGCTGGTTC
- 3′ sequencing primer CCACATAGCGTAAAAGGAGCAAC (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byThe backbone of this plasmid was derived from Addgene plasmid 22217 (from the Boyden Lab at MIT), cut with BamHI and EcoRI. The Cox8 mitochondrial targeting sequence was amplified from Addgene plasmid 23348, mito-PAGFP). The gene, ratiometric pHluorin, was originally obtained from Gero Miesenbock, Memorial Sloan-Kettering Cancer Center, New York in the pGEX2T vector.
Terms and Licenses
Ratiometric pHluorin was published in:
Miesenbock et al. “Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins” Nature 394, 192-195 (9 July 1998) | doi:10.1038/28190
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:VV064: 1xCox8 - ratiometric pHluorin in fck was a gift from Adam Cohen (Addgene plasmid # 58502 ; http://n2t.net/addgene:58502 ; RRID:Addgene_58502)
For your References section:Imaging GFP-Based Reporters in Neurons with Multiwavelength Optogenetic Control. Venkatachalam V, Cohen AE. Biophys J. 2014 Oct 7;107(7):1554-63. doi: 10.1016/j.bpj.2014.08.020. 10.1016/j.bpj.2014.08.020 PubMed 25296307