|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||25867||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4000
Vector typeMammalian Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin and Tetracycline
Growth instructionsPlasmid contains SupF. Grow in TOP10/P3 cells.
Gene/Insert nameGq alpha
Alt nameG protein alpha subunit
SpeciesM. musculus (mouse)
MutationReplacement of the five carboxyl-terminal amino acids of Gq alpha (EYNLV) with those of Gz alpha(YIGLC)
Entrez GeneGnaq (a.k.a. 1110005L02Rik, 6230401I02Rik, AA408290, AW060788, Dsk, Dsk1, Dsk10, Gal, Galphaq, Gq, GqI)
/ Fusion Protein
- HA (see comments)
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (unknown if destroyed)
- 3′ cloning site NsiI (unknown if destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer SP6 (Common Sequencing Primers)
This is Gq alpha with the C-terminal amino acids changed from Gq alpha to Gz alpha residues (EYNLV to YIGLC). Works the same as qi5 and is the least popular since no one knows what Gz alpha really does in nature. Since qz5 is not sensitive to pertussis toxin, qz5 may be the only G protein activated by a Gi-coupled receptor in cells treated with pertussis toxin. This trick can be experimentally useful in settings where you want experimental control of the exact G protein and the receptor that is activated. It is theoretically possible that this construct will work better than the other constructs for particular receptors, but the Conklin lab has not seen this happen yet.
Contains an internal hemaglutinin epitope tag (DVPDYA) that has no effect on receptor coupling.
Please see supplemental document for G-protein chimera user manual.
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:qz5 was a gift from Bruce Conklin (Addgene plasmid # 25867 ; http://n2t.net/addgene:25867 ; RRID:Addgene_25867)
For your References section:Substitution of three amino acids switches receptor specificity of Gq alpha to that of Gi alpha. Conklin BR, Farfel Z, Lustig KD, Julius D, Bourne HR. Nature. 1993 May 20. 363(6426):274-6. 10.1038/363274a0 PubMed 8387644