Purposea genetically-encoded sensor for lactate concentration, transport and metabolic flux
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||44238||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5418
- Total vector size (bp) 7648
Vector typeMammalian Expression ; lactate biosensor
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameLaconic (LACtate Optical Nano Indicator from CECs)
Insert Size (bp)2230
- Promoter CMV
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (unknown if destroyed)
- 3′ cloning site HindIII (unknown if destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer BGH-rev (Common Sequencing Primers)
Laconic is a genetically-encoded biosensor that detects lactate in the physiological range. The sensor was based on LldR, a bacterial transcription regulator that consists of a lactate-binding/regulatory domain and a DNA-binding domain. Laconic consists of a N-terminal mTFP, followed by the LldR flanked by linkers, and finally a C-terminal Venus.
See Addgene plasmid# 46307: NLS-Laconic/pCMV-myc-nuc for a nuclear-targeted version of Laconic: http://www.addgene.org/46307/ .
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:Laconic/pcDNA3.1(-) was a gift from Luis Felipe Barros (Addgene plasmid # 44238 ; http://n2t.net/addgene:44238 ; RRID:Addgene_44238)
For your References section:A Genetically Encoded FRET Lactate Sensor and Its Use To Detect the Warburg Effect in Single Cancer Cells. San_Martin A, Ceballo S, Ruminot I, Lerchundi R, Frommer WB, Barros LF. PLoS One. 2013;8(2):e57712. doi: 10.1371/journal.pone.0057712. Epub 2013 Feb 26. 10.1371/journal.pone.0057712 PubMed 23469056